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2001
Sauter C, Otalora F, Gavira J A, Vidal O, Giege R, Garcia-Ruiz J M
Structure of tetragonal hen egg-white lysozyme at 0.94 A from crystals grown by the counter-diffusion method Article de journal
Dans: Acta Crystallogr D Biol Crystallogr, vol. 57, no. Pt 8, p. 1119-1126, 2001, ISBN: 11468395, (0907-4449 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Amino Acids/chemistry Animals Anions/chemistry Binding Sites Cations/chemistry Chickens Crystallization Crystallography, Molecular Muramidase/*chemistry Protein Conformation Protein Structure, Non-U.S. Gov't, Tertiary Support, Unité ARN, X-Ray Egg White/analysis Models
@article{,
title = {Structure of tetragonal hen egg-white lysozyme at 0.94 A from crystals grown by the counter-diffusion method},
author = {C Sauter and F Otalora and J A Gavira and O Vidal and R Giege and J M Garcia-Ruiz},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11468395},
isbn = {11468395},
year = {2001},
date = {2001-01-01},
journal = {Acta Crystallogr D Biol Crystallogr},
volume = {57},
number = {Pt 8},
pages = {1119-1126},
abstract = {Very high quality crystals of tetragonal hen egg-white lysozyme were grown in the Advanced Protein Crystallization Facility (APCF) on board the Space Shuttle using a modified free-interface diffusion (FID) reactor designed ad hoc to have a longer diffusion path. This design allows the performance of true counter-diffusion experiments. Crystals were obtained under the classical chemical conditions defined 50 y ago with NaCl as a crystallizing agent and acetate pH 4.5 as a buffer. Counter-diffusion crystallization allows a "physical" instead of chemical optimization of growth conditions: indeed, this method screens for the best supersaturation conditions in a single trial and yields crystals of very high quality. A complete diffraction data set was collected at atomic resolution from one of these crystals using synchrotron radiation at the DESY-EMBL beamlines. The overall R(merge) on intensities in the resolution range 31-0.94 A was 5.2% and the data were 98.9% complete. Refinement was carried out with the programs CNS and SHELX97 to a final crystallographic R factor of 12.26% for 72 390 reflections. A mean standard uncertainty in the atomic positions of 0.024 A was estimated from inversion of blocked least-squares matrices. 22 side chains show alternate conformations and the loop 59-75 adopts in the same crystal packing two conformations that were observed for either triclinic or tetragonal lysozyme in previous high-resolution studies. In addition to 255 water molecules, the crystallizing agent (one hexacoordinated sodium ion and five chloride anions) participates in the ordered lysozyme hydration shell.},
note = {0907-4449
Journal Article},
keywords = {Amino Acids/chemistry Animals Anions/chemistry Binding Sites Cations/chemistry Chickens Crystallization Crystallography, Molecular Muramidase/*chemistry Protein Conformation Protein Structure, Non-U.S. Gov't, Tertiary Support, Unité ARN, X-Ray Egg White/analysis Models},
pubstate = {published},
tppubtype = {article}
}
Very high quality crystals of tetragonal hen egg-white lysozyme were grown in the Advanced Protein Crystallization Facility (APCF) on board the Space Shuttle using a modified free-interface diffusion (FID) reactor designed ad hoc to have a longer diffusion path. This design allows the performance of true counter-diffusion experiments. Crystals were obtained under the classical chemical conditions defined 50 y ago with NaCl as a crystallizing agent and acetate pH 4.5 as a buffer. Counter-diffusion crystallization allows a "physical" instead of chemical optimization of growth conditions: indeed, this method screens for the best supersaturation conditions in a single trial and yields crystals of very high quality. A complete diffraction data set was collected at atomic resolution from one of these crystals using synchrotron radiation at the DESY-EMBL beamlines. The overall R(merge) on intensities in the resolution range 31-0.94 A was 5.2% and the data were 98.9% complete. Refinement was carried out with the programs CNS and SHELX97 to a final crystallographic R factor of 12.26% for 72 390 reflections. A mean standard uncertainty in the atomic positions of 0.024 A was estimated from inversion of blocked least-squares matrices. 22 side chains show alternate conformations and the loop 59-75 adopts in the same crystal packing two conformations that were observed for either triclinic or tetragonal lysozyme in previous high-resolution studies. In addition to 255 water molecules, the crystallizing agent (one hexacoordinated sodium ion and five chloride anions) participates in the ordered lysozyme hydration shell.
2000
Tzou P, Ohresser S, Ferrandon Dominique, Capovilla Maria, Reichhart Jean-Marc, Lemaitre Bruno, Hoffmann Jules A, Imler Jean-Luc
Tissue-specific inducible expression of antimicrobial peptide genes in Drosophila surface epithelia Article de journal
Dans: Immunity, vol. 13, p. 737–48., 2000, ISSN: 1074-7613.
Résumé | BibTeX | Étiquettes: *Genes, Animal, Anti-Infective Agents/*immunology/metabolism, Drosophila/genetics/*immunology, ferrandon, Gene Expression Regulation/*immunology, Genes, Glycoside Hydrolases/immunology, hoffmann, Human, imler, Insect, Insect Proteins/genetics/immunology, M3i, Non-U.S. Gov't, Organ Specificity, P.H.S., reichhart, Reporter, Support, Transfection, U.S. Gov't
@article{tzou_tissue-specific_2000b,
title = {Tissue-specific inducible expression of antimicrobial peptide genes in Drosophila surface epithelia},
author = {P Tzou and S Ohresser and Dominique Ferrandon and Maria Capovilla and Jean-Marc Reichhart and Bruno Lemaitre and Jules A Hoffmann and Jean-Luc Imler},
issn = {1074-7613},
year = {2000},
date = {2000-01-01},
journal = {Immunity},
volume = {13},
pages = {737--48.},
abstract = {The production of antimicrobial peptides is an important aspect of host defense in multicellular organisms. In Drosophila, seven antimicrobial peptides with different spectra of activities are synthesized by the fat body during the immune response and secreted into the hemolymph. Using GFP reporter transgenes, we show here that all seven Drosophila antimicrobial peptides can be induced in surface epithelia in a tissue-specific manner. The imd gene plays a critical role in the activation of this local response to infection. In particular, drosomycin expression, which is regulated by the Toll pathway during the systemic response, is regulated by imd in the respiratory tract, thus demonstrating the existence of distinct regulatory mechanisms for local and systemic induction of antimicrobial peptide genes in Drosophila.},
keywords = {*Genes, Animal, Anti-Infective Agents/*immunology/metabolism, Drosophila/genetics/*immunology, ferrandon, Gene Expression Regulation/*immunology, Genes, Glycoside Hydrolases/immunology, hoffmann, Human, imler, Insect, Insect Proteins/genetics/immunology, M3i, Non-U.S. Gov't, Organ Specificity, P.H.S., reichhart, Reporter, Support, Transfection, U.S. Gov't},
pubstate = {published},
tppubtype = {article}
}
The production of antimicrobial peptides is an important aspect of host defense in multicellular organisms. In Drosophila, seven antimicrobial peptides with different spectra of activities are synthesized by the fat body during the immune response and secreted into the hemolymph. Using GFP reporter transgenes, we show here that all seven Drosophila antimicrobial peptides can be induced in surface epithelia in a tissue-specific manner. The imd gene plays a critical role in the activation of this local response to infection. In particular, drosomycin expression, which is regulated by the Toll pathway during the systemic response, is regulated by imd in the respiratory tract, thus demonstrating the existence of distinct regulatory mechanisms for local and systemic induction of antimicrobial peptide genes in Drosophila.
Rihn B H, Bottin M C, Coulais C, Rouget R, Monhoven N, Baranowski W, Edorh A, Keith G
Genotoxicity of 3-methylcholanthrene in liver of transgenic big Blue mice Article de journal
Dans: Environ Mol Mutagen, vol. 36, no. 4, p. 266-73, 2000, ISBN: 11152559, (0893-6692 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Animals Bacterial Proteins/genetics Base Sequence Cell Division/drug effects DNA Adducts DNA Primers *Escherichia coli Proteins Liver/cytology/*drug effects Methylcholanthrene/*toxicity Mice Mice, Inbred C57BL Mice, Non-U.S. Gov't, Transgenic Mutagens/*toxicity Mutation Organ Weight Repressor Proteins/genetics Support
@article{,
title = {Genotoxicity of 3-methylcholanthrene in liver of transgenic big Blue mice},
author = {B H Rihn and M C Bottin and C Coulais and R Rouget and N Monhoven and W Baranowski and A Edorh and G Keith},
editor = {Editor},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11152559},
isbn = {11152559},
year = {2000},
date = {2000-01-01},
journal = {Environ Mol Mutagen},
volume = {36},
number = {4},
pages = {266-73},
abstract = {Transgenic mice provide a unique tool for studying the tissue specificity and mutagenic potential of chemicals. Because 3-methylcholanthrene (3MC) was found mutagenic in bacteria, clastogenic in bone marrow, and induces DNA adducts in animals, we were interested to determinine whether this xenobiotic provokes (1) cell proliferation, (2) transcriptional activity changes, (3) DNA adducts, and (4) hepatic mutations in transgenic Big Blue mice carrying the lambdaLIZ phage shuttle vector. Big Blue C57/Bl male mice were treated with a single intraperitoneal dose of 80 mg/kg 3MC for 1, 3, 6, 14, or 30 days. Cell proliferation was checked by 5-bromo-2-deoxyuridine labeling and immunohistochemical detection. The maximal increase of the mitotic index was evidenced after 3 days (2.9 times the control value; P < 0.01). The relative nucleus area, reflecting the transcriptional activity, was also the highest in the treated group after 3 days: 1.86 times the control value, on average (P < 0.01). Four major DNA adducts, determined according to the [(32)P]-postlabeling method, were evidenced in liver DNA of treated mice, 6 days after the treatment: the spot intensities increased in a time-dependent manner. The mutant frequency of liver DNA was the highest after 14 days: 20.3 +/- 2.9 x 10(-5) in the treated vs. 7.6 +/- 2.7 x 10(-5) in the control mice (P < 0.01). Sequencing of the lambda lacI mutant plaques showed mainly G:C --> T:A and C:G --> A:T transversions. In conclusion, 3MC at first induced nuclear enlargement and a slight increase of cell proliferation in liver, followed by parallel formation of DNA adducts and mutations. This study shows how transgenic models allow in vivo evaluation of mechanistically simultaneous endpoints.},
note = {0893-6692
Journal Article},
keywords = {Animals Bacterial Proteins/genetics Base Sequence Cell Division/drug effects DNA Adducts DNA Primers *Escherichia coli Proteins Liver/cytology/*drug effects Methylcholanthrene/*toxicity Mice Mice, Inbred C57BL Mice, Non-U.S. Gov't, Transgenic Mutagens/*toxicity Mutation Organ Weight Repressor Proteins/genetics Support},
pubstate = {published},
tppubtype = {article}
}
Transgenic mice provide a unique tool for studying the tissue specificity and mutagenic potential of chemicals. Because 3-methylcholanthrene (3MC) was found mutagenic in bacteria, clastogenic in bone marrow, and induces DNA adducts in animals, we were interested to determinine whether this xenobiotic provokes (1) cell proliferation, (2) transcriptional activity changes, (3) DNA adducts, and (4) hepatic mutations in transgenic Big Blue mice carrying the lambdaLIZ phage shuttle vector. Big Blue C57/Bl male mice were treated with a single intraperitoneal dose of 80 mg/kg 3MC for 1, 3, 6, 14, or 30 days. Cell proliferation was checked by 5-bromo-2-deoxyuridine labeling and immunohistochemical detection. The maximal increase of the mitotic index was evidenced after 3 days (2.9 times the control value; P < 0.01). The relative nucleus area, reflecting the transcriptional activity, was also the highest in the treated group after 3 days: 1.86 times the control value, on average (P < 0.01). Four major DNA adducts, determined according to the [(32)P]-postlabeling method, were evidenced in liver DNA of treated mice, 6 days after the treatment: the spot intensities increased in a time-dependent manner. The mutant frequency of liver DNA was the highest after 14 days: 20.3 +/- 2.9 x 10(-5) in the treated vs. 7.6 +/- 2.7 x 10(-5) in the control mice (P < 0.01). Sequencing of the lambda lacI mutant plaques showed mainly G:C --> T:A and C:G --> A:T transversions. In conclusion, 3MC at first induced nuclear enlargement and a slight increase of cell proliferation in liver, followed by parallel formation of DNA adducts and mutations. This study shows how transgenic models allow in vivo evaluation of mechanistically simultaneous endpoints.
Westhof E, Fritsch V
RNA folding: beyond Watson-Crick pairs Article de journal
Dans: Structure, vol. 8, no. 3, p. R55-65, 2000, ISBN: 10745012, (0969-2126 Journal Article Review Review, Tutorial).
Résumé | Liens | BibTeX | Étiquettes: Base Pairing Human Hydrogen Bonding *Nucleic Acid Conformation RNA/*chemistry Support, Non-U.S. Gov't, Unité ARN
@article{,
title = {RNA folding: beyond Watson-Crick pairs},
author = {E Westhof and V Fritsch},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10745012},
isbn = {10745012},
year = {2000},
date = {2000-01-01},
journal = {Structure},
volume = {8},
number = {3},
pages = {R55-65},
abstract = {Several crystal structures of RNA fragments, alone or in complex with a specific protein, have been recently solved. In addition, the structures of an artificial ribozyme, the leadzyme, and the cleavage product of a human pathogen ribozyme, have extended the structural diversity of ribozyme architectures. The attained set of folding rules and motifs expand the repertoire seen previously in tRNA structures.},
note = {0969-2126
Journal Article
Review
Review, Tutorial},
keywords = {Base Pairing Human Hydrogen Bonding *Nucleic Acid Conformation RNA/*chemistry Support, Non-U.S. Gov't, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Several crystal structures of RNA fragments, alone or in complex with a specific protein, have been recently solved. In addition, the structures of an artificial ribozyme, the leadzyme, and the cleavage product of a human pathogen ribozyme, have extended the structural diversity of ribozyme architectures. The attained set of folding rules and motifs expand the repertoire seen previously in tRNA structures.
Rihn B H, Bottin M C, Coulais C, Rouget R, Monhoven N, Baranowski W, Edorh A, Keith G
Genotoxicity of 3-methylcholanthrene in liver of transgenic big Blue mice Article de journal
Dans: Environ Mol Mutagen, vol. 36, no. 4, p. 266-273, 2000, ISBN: 11152559, (0893-6692 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Animals Bacterial Proteins/genetics Base Sequence Cell Division/drug effects DNA Adducts DNA Primers *Escherichia coli Proteins Liver/cytology/*drug effects Methylcholanthrene/*toxicity Mice Mice, Inbred C57BL Mice, Non-U.S. Gov't, Transgenic Mutagens/*toxicity Mutation Organ Weight Repressor Proteins/genetics Support, Unité ARN
@article{,
title = {Genotoxicity of 3-methylcholanthrene in liver of transgenic big Blue mice},
author = {B H Rihn and M C Bottin and C Coulais and R Rouget and N Monhoven and W Baranowski and A Edorh and G Keith},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11152559},
isbn = {11152559},
year = {2000},
date = {2000-01-01},
journal = {Environ Mol Mutagen},
volume = {36},
number = {4},
pages = {266-273},
abstract = {Transgenic mice provide a unique tool for studying the tissue specificity and mutagenic potential of chemicals. Because 3-methylcholanthrene (3MC) was found mutagenic in bacteria, clastogenic in bone marrow, and induces DNA adducts in animals, we were interested to determinine whether this xenobiotic provokes (1) cell proliferation, (2) transcriptional activity changes, (3) DNA adducts, and (4) hepatic mutations in transgenic Big Blue mice carrying the lambdaLIZ phage shuttle vector. Big Blue C57/Bl male mice were treated with a single intraperitoneal dose of 80 mg/kg 3MC for 1, 3, 6, 14, or 30 days. Cell proliferation was checked by 5-bromo-2-deoxyuridine labeling and immunohistochemical detection. The maximal increase of the mitotic index was evidenced after 3 days (2.9 times the control value; P < 0.01). The relative nucleus area, reflecting the transcriptional activity, was also the highest in the treated group after 3 days: 1.86 times the control value, on average (P < 0.01). Four major DNA adducts, determined according to the [(32)P]-postlabeling method, were evidenced in liver DNA of treated mice, 6 days after the treatment: the spot intensities increased in a time-dependent manner. The mutant frequency of liver DNA was the highest after 14 days: 20.3 +/- 2.9 x 10(-5) in the treated vs. 7.6 +/- 2.7 x 10(-5) in the control mice (P < 0.01). Sequencing of the lambda lacI mutant plaques showed mainly G:C --> T:A and C:G --> A:T transversions. In conclusion, 3MC at first induced nuclear enlargement and a slight increase of cell proliferation in liver, followed by parallel formation of DNA adducts and mutations. This study shows how transgenic models allow in vivo evaluation of mechanistically simultaneous endpoints.},
note = {0893-6692
Journal Article},
keywords = {Animals Bacterial Proteins/genetics Base Sequence Cell Division/drug effects DNA Adducts DNA Primers *Escherichia coli Proteins Liver/cytology/*drug effects Methylcholanthrene/*toxicity Mice Mice, Inbred C57BL Mice, Non-U.S. Gov't, Transgenic Mutagens/*toxicity Mutation Organ Weight Repressor Proteins/genetics Support, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Transgenic mice provide a unique tool for studying the tissue specificity and mutagenic potential of chemicals. Because 3-methylcholanthrene (3MC) was found mutagenic in bacteria, clastogenic in bone marrow, and induces DNA adducts in animals, we were interested to determinine whether this xenobiotic provokes (1) cell proliferation, (2) transcriptional activity changes, (3) DNA adducts, and (4) hepatic mutations in transgenic Big Blue mice carrying the lambdaLIZ phage shuttle vector. Big Blue C57/Bl male mice were treated with a single intraperitoneal dose of 80 mg/kg 3MC for 1, 3, 6, 14, or 30 days. Cell proliferation was checked by 5-bromo-2-deoxyuridine labeling and immunohistochemical detection. The maximal increase of the mitotic index was evidenced after 3 days (2.9 times the control value; P < 0.01). The relative nucleus area, reflecting the transcriptional activity, was also the highest in the treated group after 3 days: 1.86 times the control value, on average (P < 0.01). Four major DNA adducts, determined according to the [(32)P]-postlabeling method, were evidenced in liver DNA of treated mice, 6 days after the treatment: the spot intensities increased in a time-dependent manner. The mutant frequency of liver DNA was the highest after 14 days: 20.3 +/- 2.9 x 10(-5) in the treated vs. 7.6 +/- 2.7 x 10(-5) in the control mice (P < 0.01). Sequencing of the lambda lacI mutant plaques showed mainly G:C --> T:A and C:G --> A:T transversions. In conclusion, 3MC at first induced nuclear enlargement and a slight increase of cell proliferation in liver, followed by parallel formation of DNA adducts and mutations. This study shows how transgenic models allow in vivo evaluation of mechanistically simultaneous endpoints.
Rihn B, Coulais C, Kauffer E, Bottin M C, Martin P, Yvon F, Vigneron J C, Binet S, Monhoven N, Steiblen G, Keith G
Inhaled crocidolite mutagenicity in lung DNA Article de journal
Dans: Environ Health Perspect, vol. 108, no. 4, p. 341-346, 2000, ISBN: 10753093, (0091-6765 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Air Pollutants/*adverse effects Animals Asbestos, Alveolar/physiology Male Mice Mice, Crocidolite/administration & dosage/*adverse effects DNA Adducts/*genetics DNA Damage/*genetics Inhalation Exposure Lung/*drug effects/pathology Macrophages, Non-U.S. Gov't, Transgenic Mutagenicity Tests Support, Unité ARN
@article{,
title = {Inhaled crocidolite mutagenicity in lung DNA},
author = {B Rihn and C Coulais and E Kauffer and M C Bottin and P Martin and F Yvon and J C Vigneron and S Binet and N Monhoven and G Steiblen and G Keith},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10753093},
isbn = {10753093},
year = {2000},
date = {2000-01-01},
journal = {Environ Health Perspect},
volume = {108},
number = {4},
pages = {341-346},
abstract = {We used transgenic mice carrying the lacI reporter gene to study the mutagenesis potential of asbestos crocidolite. The animals were exposed by nose-only inhalation to an aerosol containing 5.75 mg/m(3) crocidolite dust for 6 hr/day and 5 consecutive days. After 1, 4, and 12 weeks, we examined four end points: the cytology of bronchoalveolar lavage, the lung load of crocidolite, the hydrophobic DNA adducts, and the mutations in the lacI reporter gene. Twelve weeks after exposure, nearly 10% of the inhaled fibers remained in the lung (227 +/- 103 ng/mg lung). There was evidence of a typical inflammatory response consisting of multinucleate macrophages at weeks 4 and 12, whereas immediately after the exposure, we observed numerous polymorphonuclear neutrophils. The mutant frequency significatively increased during the fourth week after the exposure: 13.5 [time] 10(-5) in the exposed group versus 6. 9 10(-5) in the control group. The induction factor, defined by the ratio of checked mutants of exposed mice to checked mutants of control mice, was 1.96. The mutation spectrum of control lung DNA and exposed lung DNA was similar, suggesting the possible involvement of a DNA repair decrease in crocidolite-treated animals. We used the (32)P-postlabeling method and did not detect any increase of either 5 mC or bulky adduct in treated mice. This is the first study that demonstrates asbestos mutagenicity in vivo after a nose-only inhalation.},
note = {0091-6765
Journal Article},
keywords = {Air Pollutants/*adverse effects Animals Asbestos, Alveolar/physiology Male Mice Mice, Crocidolite/administration & dosage/*adverse effects DNA Adducts/*genetics DNA Damage/*genetics Inhalation Exposure Lung/*drug effects/pathology Macrophages, Non-U.S. Gov't, Transgenic Mutagenicity Tests Support, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
We used transgenic mice carrying the lacI reporter gene to study the mutagenesis potential of asbestos crocidolite. The animals were exposed by nose-only inhalation to an aerosol containing 5.75 mg/m(3) crocidolite dust for 6 hr/day and 5 consecutive days. After 1, 4, and 12 weeks, we examined four end points: the cytology of bronchoalveolar lavage, the lung load of crocidolite, the hydrophobic DNA adducts, and the mutations in the lacI reporter gene. Twelve weeks after exposure, nearly 10% of the inhaled fibers remained in the lung (227 +/- 103 ng/mg lung). There was evidence of a typical inflammatory response consisting of multinucleate macrophages at weeks 4 and 12, whereas immediately after the exposure, we observed numerous polymorphonuclear neutrophils. The mutant frequency significatively increased during the fourth week after the exposure: 13.5 [time] 10(-5) in the exposed group versus 6. 9 10(-5) in the control group. The induction factor, defined by the ratio of checked mutants of exposed mice to checked mutants of control mice, was 1.96. The mutation spectrum of control lung DNA and exposed lung DNA was similar, suggesting the possible involvement of a DNA repair decrease in crocidolite-treated animals. We used the (32)P-postlabeling method and did not detect any increase of either 5 mC or bulky adduct in treated mice. This is the first study that demonstrates asbestos mutagenicity in vivo after a nose-only inhalation.
Masquida B, Westhof E
On the wobble GoU and related pairs Article de journal
Dans: RNA, vol. 6, no. 1, p. 9-15, 2000, ISBN: 10668794, (1355-8382 Journal Article Review Review, Tutorial).
Résumé | Liens | BibTeX | Étiquettes: *Base Pairing Models, Molecular RNA/*chemistry Support, Non-U.S. Gov't, Unité ARN
@article{,
title = {On the wobble GoU and related pairs},
author = {B Masquida and E Westhof},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10668794},
isbn = {10668794},
year = {2000},
date = {2000-01-01},
journal = {RNA},
volume = {6},
number = {1},
pages = {9-15},
abstract = {The wobble GoU pairs have been implicated in several biological processes where RNA molecules play a key role. We review the geometrical and conformational properties of wobble GoU pairs on the basis of available crystal structures of RNAs at high resolution. The similarities with the wobble A+oC pairs and UoU pairs are illustrated, while the differences with the recently discovered bifurcated G x U pairs are contrasted.},
note = {1355-8382
Journal Article
Review
Review, Tutorial},
keywords = {*Base Pairing Models, Molecular RNA/*chemistry Support, Non-U.S. Gov't, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
The wobble GoU pairs have been implicated in several biological processes where RNA molecules play a key role. We review the geometrical and conformational properties of wobble GoU pairs on the basis of available crystal structures of RNAs at high resolution. The similarities with the wobble A+oC pairs and UoU pairs are illustrated, while the differences with the recently discovered bifurcated G x U pairs are contrasted.
Lodmell J S, Ehresmann C, Ehresmann B, Marquet R
Convergence of natural and artificial evolution on an RNA loop-loop interaction: the HIV-1 dimerization initiation site Article de journal
Dans: RNA, vol. 6, no. 9, p. 1267-1276, 2000, ISBN: 10999604, (1355-8382 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Codon, Initiator Dimerization Directed Molecular Evolution Evolution, MARQUET, Molecular HIV-1/*chemistry/genetics Nucleic Acid Conformation RNA, Non-U.S. Gov't, Unité ARN, Viral/*chemistry/metabolism Support
@article{,
title = {Convergence of natural and artificial evolution on an RNA loop-loop interaction: the HIV-1 dimerization initiation site},
author = {J S Lodmell and C Ehresmann and B Ehresmann and R Marquet},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10999604},
isbn = {10999604},
year = {2000},
date = {2000-01-01},
journal = {RNA},
volume = {6},
number = {9},
pages = {1267-1276},
abstract = {Loop-loop interactions among nucleic acids constitute an important form of molecular recognition in a variety of biological systems. In HIV-1, genomic dimerization involves an intermolecular RNA loop-loop interaction at the dimerization initiation site (DIS), a hairpin located in the 5' noncoding region that contains an autocomplementary sequence in the loop. Only two major DIS loop sequence variants are observed among natural viral isolates. To investigate sequence and structural constraints on genomic RNA dimerization as well as loop-loop interactions in general, we randomized several or all of the nucleotides in the DIS loop and selected in vitro for dimerization-competent sequences. Surprisingly, increasing interloop complementarity above a threshold of 6 bp did not enhance dimerization, although the combinations of nucleotides forming the theoretically most stable hexanucleotide duplexes were selected. Noncanonical interactions contributed significantly to the stability and/or specificity of the dimeric complexes as demonstrated by the overwhelming bias for noncanonical base pairs closing the loop and covariations between flanking and central loop nucleotides. Degeneration of the entire loop yielded a complex population of dimerization-competent sequences whose consensus sequence resembles that of wild-type HIV-1. We conclude from these findings that the DIS has evolved to satisfy simultaneous constraints for optimal dimerization affinity and the capacity for homodimerization. Furthermore, the most constrained features of the DIS identified by our experiments could be the basis for the rational design of DIS-targeted antiviral compounds.},
note = {1355-8382
Journal Article},
keywords = {Codon, Initiator Dimerization Directed Molecular Evolution Evolution, MARQUET, Molecular HIV-1/*chemistry/genetics Nucleic Acid Conformation RNA, Non-U.S. Gov't, Unité ARN, Viral/*chemistry/metabolism Support},
pubstate = {published},
tppubtype = {article}
}
Loop-loop interactions among nucleic acids constitute an important form of molecular recognition in a variety of biological systems. In HIV-1, genomic dimerization involves an intermolecular RNA loop-loop interaction at the dimerization initiation site (DIS), a hairpin located in the 5' noncoding region that contains an autocomplementary sequence in the loop. Only two major DIS loop sequence variants are observed among natural viral isolates. To investigate sequence and structural constraints on genomic RNA dimerization as well as loop-loop interactions in general, we randomized several or all of the nucleotides in the DIS loop and selected in vitro for dimerization-competent sequences. Surprisingly, increasing interloop complementarity above a threshold of 6 bp did not enhance dimerization, although the combinations of nucleotides forming the theoretically most stable hexanucleotide duplexes were selected. Noncanonical interactions contributed significantly to the stability and/or specificity of the dimeric complexes as demonstrated by the overwhelming bias for noncanonical base pairs closing the loop and covariations between flanking and central loop nucleotides. Degeneration of the entire loop yielded a complex population of dimerization-competent sequences whose consensus sequence resembles that of wild-type HIV-1. We conclude from these findings that the DIS has evolved to satisfy simultaneous constraints for optimal dimerization affinity and the capacity for homodimerization. Furthermore, the most constrained features of the DIS identified by our experiments could be the basis for the rational design of DIS-targeted antiviral compounds.
Kolb F A, Malmgren C, Westhof E, Ehresmann C, Ehresmann B, Wagner E G, Romby P
An unusual structure formed by antisense-target RNA binding involves an extended kissing complex with a four-way junction and a side-by-side helical alignment Article de journal
Dans: RNA, vol. 6, no. 3, p. 311-324, 2000, ISBN: 10744017, (1355-8382 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Antisense/*metabolism RNA, Bacterial Proteins/*metabolism Base Pairing Base Sequence Binding Sites Cations, Divalent Computer Simulation Metals, Double-Stranded/metabolism RNA, Heavy/metabolism Models, Messenger/metabolism RNA, Molecular Molecular Sequence Data *Nucleic Acid Conformation RNA Stability RNA, Non-U.S. Gov't, ROMBY, Spliced Leader/metabolism Support, Unité ARN
@article{,
title = {An unusual structure formed by antisense-target RNA binding involves an extended kissing complex with a four-way junction and a side-by-side helical alignment},
author = {F A Kolb and C Malmgren and E Westhof and C Ehresmann and B Ehresmann and E G Wagner and P Romby},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10744017},
isbn = {10744017},
year = {2000},
date = {2000-01-01},
journal = {RNA},
volume = {6},
number = {3},
pages = {311-324},
abstract = {The antisense RNA CopA binds to the leader region of the repA mRNA (target: CopT). Previous studies on CopA-CopT pairing in vitro showed that the dominant product of antisense RNA-mRNA binding is not a full RNA duplex. We have studied here the structure of CopA-CopT complex, combining chemical and enzymatic probing and computer graphic modeling. CopI, a truncated derivative of CopA unable to bind CopT stably, was also analyzed. We show here that after initial loop-loop interaction (kissing), helix propagation resulted in an extended kissing complex that involves the formation of two intermolecular helices. By introducing mutations (base-pair inversions) into the upper stem regions of CopA and CopT, the boundaries of the two newly formed intermolecular helices were delimited. The resulting extended kissing complex represents a new type of four-way junction structure that adopts an asymmetrical X-shaped conformation formed by two helical domains, each one generated by coaxial stacking of two helices. This structure motif induces a side-by-side alignment of two long intramolecular helices that, in turn, facilitates the formation of an additional intermolecular helix that greatly stabilizes the inhibitory CopA-CopT RNA complex. This stabilizer helix cannot form in CopI-CopT complexes due to absence of the sequences involved. The functional significance of the three-dimensional models of the extended kissing complex (CopI-CopT) and the stable complex (CopA-CopT) are discussed.},
note = {1355-8382
Journal Article},
keywords = {Antisense/*metabolism RNA, Bacterial Proteins/*metabolism Base Pairing Base Sequence Binding Sites Cations, Divalent Computer Simulation Metals, Double-Stranded/metabolism RNA, Heavy/metabolism Models, Messenger/metabolism RNA, Molecular Molecular Sequence Data *Nucleic Acid Conformation RNA Stability RNA, Non-U.S. Gov't, ROMBY, Spliced Leader/metabolism Support, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
The antisense RNA CopA binds to the leader region of the repA mRNA (target: CopT). Previous studies on CopA-CopT pairing in vitro showed that the dominant product of antisense RNA-mRNA binding is not a full RNA duplex. We have studied here the structure of CopA-CopT complex, combining chemical and enzymatic probing and computer graphic modeling. CopI, a truncated derivative of CopA unable to bind CopT stably, was also analyzed. We show here that after initial loop-loop interaction (kissing), helix propagation resulted in an extended kissing complex that involves the formation of two intermolecular helices. By introducing mutations (base-pair inversions) into the upper stem regions of CopA and CopT, the boundaries of the two newly formed intermolecular helices were delimited. The resulting extended kissing complex represents a new type of four-way junction structure that adopts an asymmetrical X-shaped conformation formed by two helical domains, each one generated by coaxial stacking of two helices. This structure motif induces a side-by-side alignment of two long intramolecular helices that, in turn, facilitates the formation of an additional intermolecular helix that greatly stabilizes the inhibitory CopA-CopT RNA complex. This stabilizer helix cannot form in CopI-CopT complexes due to absence of the sequences involved. The functional significance of the three-dimensional models of the extended kissing complex (CopI-CopT) and the stable complex (CopA-CopT) are discussed.
Kolb F A, Engdahl H M, Slagter-Jager J G, Ehresmann B, Ehresmann C, Westhof E, Wagner E G, Romby P
Progression of a loop-loop complex to a four-way junction is crucial for the activity of a regulatory antisense RNA Article de journal
Dans: EMBO J, vol. 19, no. 21, p. 5905-5915, 2000, ISBN: 11060041, (0261-4189 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Antisense/*chemistry/*genetics/metabolism RNA, Bacterial Models, Bacterial Proteins/genetics Base Sequence Binding, Bacterial/chemistry/genetics/metabolism Support, Competitive DNA Primers/genetics Escherichia coli/chemistry/genetics/metabolism Genes, Molecular Molecular Sequence Data Mutation Nucleic Acid Conformation RNA, Non-U.S. Gov't, ROMBY, Unité ARN
@article{,
title = {Progression of a loop-loop complex to a four-way junction is crucial for the activity of a regulatory antisense RNA},
author = {F A Kolb and H M Engdahl and J G Slagter-Jager and B Ehresmann and C Ehresmann and E Westhof and E G Wagner and P Romby},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11060041},
isbn = {11060041},
year = {2000},
date = {2000-01-01},
journal = {EMBO J},
volume = {19},
number = {21},
pages = {5905-5915},
abstract = {The antisense RNA, CopA, regulates the replication frequency of plasmid R1 through inhibition of RepA translation by rapid and specific binding to its target RNA (CopT). The stable CopA-CopT complex is characterized by a four-way junction structure and a side-by-side alignment of two long intramolecular helices. The significance of this structure for binding in vitro and control in vivo was tested by mutations in both CopA and CopT. High rates of stable complex formation in vitro and efficient inhibition in vivo required initial loop-loop complexes to be rapidly converted to extended interactions. These interactions involve asymmetric helix progression and melting of the upper stems of both RNAs to promote the formation of two intermolecular helices. Data presented here delineate the boundaries of these helices and emphasize the need for unimpeded helix propagation. This process is directional, i.e. one of the two intermolecular helices (B) must form first to allow formation of the other (B'). A binding pathway, characterized by a hierarchy of intermediates leading to an irreversible and inhibitory RNA-RNA complex, is proposed.},
note = {0261-4189
Journal Article},
keywords = {Antisense/*chemistry/*genetics/metabolism RNA, Bacterial Models, Bacterial Proteins/genetics Base Sequence Binding, Bacterial/chemistry/genetics/metabolism Support, Competitive DNA Primers/genetics Escherichia coli/chemistry/genetics/metabolism Genes, Molecular Molecular Sequence Data Mutation Nucleic Acid Conformation RNA, Non-U.S. Gov't, ROMBY, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
The antisense RNA, CopA, regulates the replication frequency of plasmid R1 through inhibition of RepA translation by rapid and specific binding to its target RNA (CopT). The stable CopA-CopT complex is characterized by a four-way junction structure and a side-by-side alignment of two long intramolecular helices. The significance of this structure for binding in vitro and control in vivo was tested by mutations in both CopA and CopT. High rates of stable complex formation in vitro and efficient inhibition in vivo required initial loop-loop complexes to be rapidly converted to extended interactions. These interactions involve asymmetric helix progression and melting of the upper stems of both RNAs to promote the formation of two intermolecular helices. Data presented here delineate the boundaries of these helices and emphasize the need for unimpeded helix propagation. This process is directional, i.e. one of the two intermolecular helices (B) must form first to allow formation of the other (B'). A binding pathway, characterized by a hierarchy of intermediates leading to an irreversible and inhibitory RNA-RNA complex, is proposed.
Giege R, Felden B, Zenkova M A, Sil'nikov V N, Vlassov V V
Cleavage of RNA with synthetic ribonuclease mimics Article de journal
Dans: Methods Enzymol, vol. 318, p. 147-165, 2000, ISBN: 10889986, (0076-6879 Journal Article).
Liens | BibTeX | Étiquettes: Asp/chemistry Ribonuclease, Base Sequence Electrophoresis, Chemical Molecular Sequence Data Nucleic Acid Conformation Nucleic Acid Hybridization Phosphorylation Plasmids/metabolism RNA/chemistry/*metabolism RNA, Non-U.S. Gov't, Pancreatic/chemistry/pharmacology Ribonucleases/*chemistry/pharmacology Saccharomyces cerevisiae/genetics Spectrophotometry Support, Polyacrylamide Gel Genetic Techniques Hydrolysis Imidazoles/pharmacology Models, Transfer, Unité ARN
@article{,
title = {Cleavage of RNA with synthetic ribonuclease mimics},
author = {R Giege and B Felden and M A Zenkova and V N Sil'nikov and V V Vlassov},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10889986},
isbn = {10889986},
year = {2000},
date = {2000-01-01},
journal = {Methods Enzymol},
volume = {318},
pages = {147-165},
note = {0076-6879
Journal Article},
keywords = {Asp/chemistry Ribonuclease, Base Sequence Electrophoresis, Chemical Molecular Sequence Data Nucleic Acid Conformation Nucleic Acid Hybridization Phosphorylation Plasmids/metabolism RNA/chemistry/*metabolism RNA, Non-U.S. Gov't, Pancreatic/chemistry/pharmacology Ribonucleases/*chemistry/pharmacology Saccharomyces cerevisiae/genetics Spectrophotometry Support, Polyacrylamide Gel Genetic Techniques Hydrolysis Imidazoles/pharmacology Models, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Geslain R, Martin F, Delagoutte B, Cavarelli J, Gangloff J, Eriani G
In vivo selection of lethal mutations reveals two functional domains in arginyl-tRNA synthetase Article de journal
Dans: RNA, vol. 6, no. 3, p. 434-448, 2000, ISBN: 10744027, (1355-8382 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Arginine-tRNA Ligase/chemistry/*genetics Cloning, ERIANI, Fungal Genes, Fungal/genetics Kinetics Models, Lethal/*genetics Genes, Molecular Fungal Proteins/biosynthesis/genetics Gene Expression Regulation, Molecular Mutation/*genetics Peptide Fragments/chemistry/genetics Saccharomyces cerevisiae/enzymology/genetics Support, Non-U.S. Gov't, Structural, Unité ARN
@article{,
title = {In vivo selection of lethal mutations reveals two functional domains in arginyl-tRNA synthetase},
author = {R Geslain and F Martin and B Delagoutte and J Cavarelli and J Gangloff and G Eriani},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10744027},
isbn = {10744027},
year = {2000},
date = {2000-01-01},
journal = {RNA},
volume = {6},
number = {3},
pages = {434-448},
abstract = {Using random mutagenesis and a genetic screening in yeast, we isolated 26 mutations that inactivate Saccharomyces cerevisiae arginyl-tRNA synthetase (ArgRS). The mutations were identified and the kinetic parameters of the corresponding proteins were tested after purification of the expression products in Escherichia coli. The effects were interpreted in the light of the crystal structure of ArgRS. Eighteen functional residues were found around the arginine-binding pocket and eight others in the carboxy-terminal domain of the enzyme. Mutations of these residues all act by strongly impairing the rates of tRNA charging and arginine activation. Thus, ArgRS and tRNA(Arg) can be considered as a kind of ribonucleoprotein, where the tRNA, before being charged, is acting as a cofactor that activates the enzyme. Furthermore, by using different tRNA(Arg) isoacceptors and heterologous tRNA(Asp), we highlighted the crucial role of several residues of the carboxy-terminal domain in tRNA recognition and discrimination.},
note = {1355-8382
Journal Article},
keywords = {Arginine-tRNA Ligase/chemistry/*genetics Cloning, ERIANI, Fungal Genes, Fungal/genetics Kinetics Models, Lethal/*genetics Genes, Molecular Fungal Proteins/biosynthesis/genetics Gene Expression Regulation, Molecular Mutation/*genetics Peptide Fragments/chemistry/genetics Saccharomyces cerevisiae/enzymology/genetics Support, Non-U.S. Gov't, Structural, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Using random mutagenesis and a genetic screening in yeast, we isolated 26 mutations that inactivate Saccharomyces cerevisiae arginyl-tRNA synthetase (ArgRS). The mutations were identified and the kinetic parameters of the corresponding proteins were tested after purification of the expression products in Escherichia coli. The effects were interpreted in the light of the crystal structure of ArgRS. Eighteen functional residues were found around the arginine-binding pocket and eight others in the carboxy-terminal domain of the enzyme. Mutations of these residues all act by strongly impairing the rates of tRNA charging and arginine activation. Thus, ArgRS and tRNA(Arg) can be considered as a kind of ribonucleoprotein, where the tRNA, before being charged, is acting as a cofactor that activates the enzyme. Furthermore, by using different tRNA(Arg) isoacceptors and heterologous tRNA(Asp), we highlighted the crucial role of several residues of the carboxy-terminal domain in tRNA recognition and discrimination.
Frugier M, Moulinier L, Giege R
A domain in the N-terminal extension of class IIb eukaryotic aminoacyl-tRNA synthetases is important for tRNA binding Article de journal
Dans: EMBO J, vol. 19, no. 10, p. 2371-2380, 2000, ISBN: 10811628, (0261-4189 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Amino Acid Sequence Amino Acyl-tRNA Ligases/chemistry/*metabolism Aspartate-tRNA Ligase/chemistry/metabolism Molecular Sequence Data RNA, ERIANI, FRUGIER, Fungal/metabolism RNA, Non-U.S. Gov't, Transfer/*metabolism Saccharomyces cerevisiae/*metabolism Sequence Alignment Support, Unité ARN
@article{,
title = {A domain in the N-terminal extension of class IIb eukaryotic aminoacyl-tRNA synthetases is important for tRNA binding},
author = {M Frugier and L Moulinier and R Giege},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10811628},
isbn = {10811628},
year = {2000},
date = {2000-01-01},
journal = {EMBO J},
volume = {19},
number = {10},
pages = {2371-2380},
abstract = {Cytoplasmic aspartyl-tRNA synthetase (AspRS) from Saccharomyces cerevisiae is a homodimer of 64 kDa subunits. Previous studies have emphasized the high sensitivity of the N-terminal region to proteolytic cleavage, leading to truncated species that have lost the first 20-70 residues but that retain enzymatic activity and dimeric structure. In this work, we demonstrate that the N-terminal extension in yeast AspRS participates in tRNA binding and we generalize this finding to eukaryotic class IIb aminoacyl-tRNA synthetases. By gel retardation studies and footprinting experiments on yeast tRNA(Asp), we show that the extension, connected to the anticodon-binding module of the synthetase, contacts tRNA on the minor groove side of its anticodon stem. Sequence comparison of eukaryotic class IIb synthetases identifies a lysine-rich 11 residue sequence ((29)LSKKALKKLQK(39) in yeast AspRS with the consensus xSKxxLKKxxK in class IIb synthetases) that is important for this binding. Direct proof of the role of this sequence comes from a mutagenesis analysis and from binding studies using the isolated peptide.},
note = {0261-4189
Journal Article},
keywords = {Amino Acid Sequence Amino Acyl-tRNA Ligases/chemistry/*metabolism Aspartate-tRNA Ligase/chemistry/metabolism Molecular Sequence Data RNA, ERIANI, FRUGIER, Fungal/metabolism RNA, Non-U.S. Gov't, Transfer/*metabolism Saccharomyces cerevisiae/*metabolism Sequence Alignment Support, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Cytoplasmic aspartyl-tRNA synthetase (AspRS) from Saccharomyces cerevisiae is a homodimer of 64 kDa subunits. Previous studies have emphasized the high sensitivity of the N-terminal region to proteolytic cleavage, leading to truncated species that have lost the first 20-70 residues but that retain enzymatic activity and dimeric structure. In this work, we demonstrate that the N-terminal extension in yeast AspRS participates in tRNA binding and we generalize this finding to eukaryotic class IIb aminoacyl-tRNA synthetases. By gel retardation studies and footprinting experiments on yeast tRNA(Asp), we show that the extension, connected to the anticodon-binding module of the synthetase, contacts tRNA on the minor groove side of its anticodon stem. Sequence comparison of eukaryotic class IIb synthetases identifies a lysine-rich 11 residue sequence ((29)LSKKALKKLQK(39) in yeast AspRS with the consensus xSKxxLKKxxK in class IIb synthetases) that is important for this binding. Direct proof of the role of this sequence comes from a mutagenesis analysis and from binding studies using the isolated peptide.
Fagegaltier D, Hubert N, Carbon P, Krol A
The selenocysteine insertion sequence binding protein SBP is different from the Y-box protein dbpB Article de journal
Dans: Biochimie, vol. 82, no. 2, p. 117-122, 2000, ISBN: 10727766, (0300-9084 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Animals Base Sequence *CCAAT-Enhancer-Binding Proteins COS Cells Cloning, Molecular DNA-Binding Proteins/genetics/*metabolism Molecular Sequence Data Nucleic Acid Conformation RNA/metabolism RNA-Binding Proteins/genetics/*metabolism Rats Support, Non-U.S. Gov't, Unité ARN
@article{,
title = {The selenocysteine insertion sequence binding protein SBP is different from the Y-box protein dbpB},
author = {D Fagegaltier and N Hubert and P Carbon and A Krol},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10727766},
isbn = {10727766},
year = {2000},
date = {2000-01-01},
journal = {Biochimie},
volume = {82},
number = {2},
pages = {117-122},
abstract = {In eukaryotes, translation of internal UGA selenocysteine codons requires the SECIS stem-loop structure in the 3'UTR of selenoprotein mRNAs. In an earlier work, we identified SBP as a selenocysteine insertion sequence (SECIS)-binding protein. Here, the yeast three-hybrid screen was employed to capture the cDNA of SBP. One candidate, satisfying the genetic screens, was identified as the already known dbpB protein. Although it was also found by another group, but with a different strategy, to carry SECIS-binding activity, further experiments enabled us to show that dbpB was unable to bind the SECIS element in vitro. Altogether, our findings led us to conclude that, under our conditions, dbpB and SBP are two distinct proteins.},
note = {0300-9084
Journal Article},
keywords = {Animals Base Sequence *CCAAT-Enhancer-Binding Proteins COS Cells Cloning, Molecular DNA-Binding Proteins/genetics/*metabolism Molecular Sequence Data Nucleic Acid Conformation RNA/metabolism RNA-Binding Proteins/genetics/*metabolism Rats Support, Non-U.S. Gov't, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
In eukaryotes, translation of internal UGA selenocysteine codons requires the SECIS stem-loop structure in the 3'UTR of selenoprotein mRNAs. In an earlier work, we identified SBP as a selenocysteine insertion sequence (SECIS)-binding protein. Here, the yeast three-hybrid screen was employed to capture the cDNA of SBP. One candidate, satisfying the genetic screens, was identified as the already known dbpB protein. Although it was also found by another group, but with a different strategy, to carry SECIS-binding activity, further experiments enabled us to show that dbpB was unable to bind the SECIS element in vitro. Altogether, our findings led us to conclude that, under our conditions, dbpB and SBP are two distinct proteins.
Delagoutte B, Keith G, Moras D, Cavarelli J
Crystallization and preliminary X-ray crystallographic analysis of yeast arginyl-tRNA synthetase-yeast tRNAArg complexes Article de journal
Dans: Acta Crystallogr D Biol Crystallogr, vol. 56, no. Pt 4, p. 492-494, 2000, ISBN: 10739930, (0907-4449 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Arg/*chemistry/isolation & purification/*metabolism Saccharomyces cerevisiae/enzymology/genetics Support, Arginine-tRNA Ligase/*chemistry/isolation & purification/*metabolism Crystallization Crystallography, Fungal/chemistry/isolation & purification/metabolism RNA, Non-U.S. Gov't, Transfer, Unité ARN, X-Ray RNA
@article{,
title = {Crystallization and preliminary X-ray crystallographic analysis of yeast arginyl-tRNA synthetase-yeast tRNAArg complexes},
author = {B Delagoutte and G Keith and D Moras and J Cavarelli},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10739930},
isbn = {10739930},
year = {2000},
date = {2000-01-01},
journal = {Acta Crystallogr D Biol Crystallogr},
volume = {56},
number = {Pt 4},
pages = {492-494},
abstract = {Three different crystal forms of complexes between arginyl-tRNA synthetase from the yeast Saccharomyces cerevisae (yArgRS) and the yeast second major tRNA(Arg) (tRNA(Arg)(ICG)) isoacceptor have been crystallized by the hanging-drop vapour-diffusion method in the presence of ammonium sulfate. Crystal form II, which diffracts beyond 2.2 A resolution at the European Synchrotron Radiation Facility ID14-4 beamline, belongs to the orthorhombic space group P2(1)2(1)2, with unit-cell parameters a = 129.64},
note = {0907-4449
Journal Article},
keywords = {Arg/*chemistry/isolation & purification/*metabolism Saccharomyces cerevisiae/enzymology/genetics Support, Arginine-tRNA Ligase/*chemistry/isolation & purification/*metabolism Crystallization Crystallography, Fungal/chemistry/isolation & purification/metabolism RNA, Non-U.S. Gov't, Transfer, Unité ARN, X-Ray RNA},
pubstate = {published},
tppubtype = {article}
}
Three different crystal forms of complexes between arginyl-tRNA synthetase from the yeast Saccharomyces cerevisae (yArgRS) and the yeast second major tRNA(Arg) (tRNA(Arg)(ICG)) isoacceptor have been crystallized by the hanging-drop vapour-diffusion method in the presence of ammonium sulfate. Crystal form II, which diffracts beyond 2.2 A resolution at the European Synchrotron Radiation Facility ID14-4 beamline, belongs to the orthorhombic space group P2(1)2(1)2, with unit-cell parameters a = 129.64
Benito Y, Kolb F A, Romby P, Lina G, Etienne J, Vandenesch F
Probing the structure of RNAIII, the Staphylococcus aureus agr regulatory RNA, and identification of the RNA domain involved in repression of protein A expression Article de journal
Dans: RNA, vol. 6, no. 5, p. 668-679, 2000, ISBN: 10836788, (1355-8382 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Antisense/*chemistry/genetics/metabolism RNA, Bacterial Models, Bacterial/*chemistry/genetics/metabolism Ribosomes/metabolism Staphylococcal Protein A/*genetics Staphylococcus aureus/*chemistry/*genetics/metabolism Support, Base Sequence Binding Sites/genetics DNA Primers/genetics Escherichia coli/metabolism Gene Expression Genes, Molecular Molecular Sequence Data Nucleic Acid Conformation RNA, Non-U.S. Gov't, ROMBY, Unité ARN
@article{,
title = {Probing the structure of RNAIII, the Staphylococcus aureus agr regulatory RNA, and identification of the RNA domain involved in repression of protein A expression},
author = {Y Benito and F A Kolb and P Romby and G Lina and J Etienne and F Vandenesch},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10836788},
isbn = {10836788},
year = {2000},
date = {2000-01-01},
journal = {RNA},
volume = {6},
number = {5},
pages = {668-679},
abstract = {RNAIII, a 514-nt RNA molecule, regulates the expression of many Staphylococcus aureus genes encoding exoproteins and cell-wall-associated proteins. We have studied the structure of RNAIII in solution, using a combination of chemical and enzymatic probes. A model of the secondary structure was derived from experimental data with the help of computer simulation of RNA folding. The model contains 14 hairpin structures connected by unpaired nucleotides. The data also point to three helices formed by distant nucleotides that close off structural domains. This model was generally compatible with the results of in vivo probing experiments with dimethylsulfate in late exponential-phase cultures. Toe-printing experiments revealed that the ribosome binding site of hld, which is encoded by RNAIII, was accessible to the Escherichia coli 30S ribosomal subunit, suggesting that the in vitro structure represented a translatable form of RNAIII. We also found that, within the 3' end of RNAIII, the conserved hairpin 13 and the terminator form an intrinsic structural domain that exerts specific regulatory activity on protein A gene expression.},
note = {1355-8382
Journal Article},
keywords = {Antisense/*chemistry/genetics/metabolism RNA, Bacterial Models, Bacterial/*chemistry/genetics/metabolism Ribosomes/metabolism Staphylococcal Protein A/*genetics Staphylococcus aureus/*chemistry/*genetics/metabolism Support, Base Sequence Binding Sites/genetics DNA Primers/genetics Escherichia coli/metabolism Gene Expression Genes, Molecular Molecular Sequence Data Nucleic Acid Conformation RNA, Non-U.S. Gov't, ROMBY, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
RNAIII, a 514-nt RNA molecule, regulates the expression of many Staphylococcus aureus genes encoding exoproteins and cell-wall-associated proteins. We have studied the structure of RNAIII in solution, using a combination of chemical and enzymatic probes. A model of the secondary structure was derived from experimental data with the help of computer simulation of RNA folding. The model contains 14 hairpin structures connected by unpaired nucleotides. The data also point to three helices formed by distant nucleotides that close off structural domains. This model was generally compatible with the results of in vivo probing experiments with dimethylsulfate in late exponential-phase cultures. Toe-printing experiments revealed that the ribosome binding site of hld, which is encoded by RNAIII, was accessible to the Escherichia coli 30S ribosomal subunit, suggesting that the in vitro structure represented a translatable form of RNAIII. We also found that, within the 3' end of RNAIII, the conserved hairpin 13 and the terminator form an intrinsic structural domain that exerts specific regulatory activity on protein A gene expression.
Benas P, Bec G, Keith G, Marquet R, Ehresmann C, Ehresmann B, Dumas P
The crystal structure of HIV reverse-transcription primer tRNA(Lys,3) shows a canonical anticodon loop Article de journal
Dans: RNA, vol. 6, no. 10, p. 1347-1355, 2000, ISBN: 11073212, (1355-8382 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Animals Anticodon/*chemistry/genetics Base Sequence Cattle Chickens/*genetics Crystallography, Biomolecular *Nucleic Acid Conformation RNA/*chemistry/genetics RNA, Lys/*chemistry/genetics Rabbits Support, MARQUET, Molecular Molecular Sequence Data Nuclear Magnetic Resonance, Non-U.S. Gov't, Transfer, Unité ARN, X-Ray HIV-1/*genetics Models
@article{,
title = {The crystal structure of HIV reverse-transcription primer tRNA(Lys,3) shows a canonical anticodon loop},
author = {P Benas and G Bec and G Keith and R Marquet and C Ehresmann and B Ehresmann and P Dumas},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11073212},
isbn = {11073212},
year = {2000},
date = {2000-01-01},
journal = {RNA},
volume = {6},
number = {10},
pages = {1347-1355},
abstract = {We have solved to 3.3 A resolution the crystal structure of the HIV reverse-transcription primer tRNA(Lys,3). The overall structure is exactly comparable to the well-known L-shape structure first revealed by yeast tRNA(Phe). In particular, it unambiguously shows a canonical anticodon loop. This contradicts previous results in short RNA fragment studies and leads us to conclude that neither frameshifting specificities of tRNA(Lys) nor tRNA(Lys,3) primer selection by HIV are due to a specific three-dimensional anticodon structure. Comparison of our structure with the results of an NMR study on a hairpin representing a nonmodified anticodon stem-loop makes plausible the conclusion that chemical modifications of the wobble base U34 to 5-methoxycarbonyl-methyl-2-thiouridine and of A37 to 2-methylthio-N-6-threonylcarbamoyl-adenosine would be responsible for a canonical 7-nt anticodon-loop structure, whereas the unmodified form would result in a noncanonical UUU short triloop. The hexagonal crystal packing is remarkable and shows tight dimers of tRNAs forming a right-handed double superhelix. Within the dimers, the tRNAs are associated head-to-tail such that the CCA end of one tRNA interacts with the anticodon of the symmetry-related tRNA. This provides us with a partial view of a codon-anticodon interaction and gives insights into the positioning of residue 37, and of its posttranscriptional modifications, relative to the first base of the codon.},
note = {1355-8382
Journal Article},
keywords = {Animals Anticodon/*chemistry/genetics Base Sequence Cattle Chickens/*genetics Crystallography, Biomolecular *Nucleic Acid Conformation RNA/*chemistry/genetics RNA, Lys/*chemistry/genetics Rabbits Support, MARQUET, Molecular Molecular Sequence Data Nuclear Magnetic Resonance, Non-U.S. Gov't, Transfer, Unité ARN, X-Ray HIV-1/*genetics Models},
pubstate = {published},
tppubtype = {article}
}
We have solved to 3.3 A resolution the crystal structure of the HIV reverse-transcription primer tRNA(Lys,3). The overall structure is exactly comparable to the well-known L-shape structure first revealed by yeast tRNA(Phe). In particular, it unambiguously shows a canonical anticodon loop. This contradicts previous results in short RNA fragment studies and leads us to conclude that neither frameshifting specificities of tRNA(Lys) nor tRNA(Lys,3) primer selection by HIV are due to a specific three-dimensional anticodon structure. Comparison of our structure with the results of an NMR study on a hairpin representing a nonmodified anticodon stem-loop makes plausible the conclusion that chemical modifications of the wobble base U34 to 5-methoxycarbonyl-methyl-2-thiouridine and of A37 to 2-methylthio-N-6-threonylcarbamoyl-adenosine would be responsible for a canonical 7-nt anticodon-loop structure, whereas the unmodified form would result in a noncanonical UUU short triloop. The hexagonal crystal packing is remarkable and shows tight dimers of tRNAs forming a right-handed double superhelix. Within the dimers, the tRNAs are associated head-to-tail such that the CCA end of one tRNA interacts with the anticodon of the symmetry-related tRNA. This provides us with a partial view of a codon-anticodon interaction and gives insights into the positioning of residue 37, and of its posttranscriptional modifications, relative to the first base of the codon.
1999
Yusupov M, Walter P, Marquet R, Ehresmann C, Ehresmann B, Dumas P
Crystallization of the dimerization-initiation site of genomic HIV-1 RNA: preliminary crystallographic results Article de journal
Dans: Acta Crystallogr D Biol Crystallogr, vol. 55, no. Pt 1, p. 281-284, 1999, ISBN: 10089425, (0907-4449 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Base Sequence Binding Sites Crystallization Crystallography, MARQUET, Non-U.S. Gov't, Unité ARN, Viral HIV-1/*chemistry/genetics Human Molecular Sequence Data Nucleic Acid Conformation RNA, Viral/*chemistry/genetics/isolation & purification Solutions Support, X-Ray Dimerization Genome
@article{,
title = {Crystallization of the dimerization-initiation site of genomic HIV-1 RNA: preliminary crystallographic results},
author = {M Yusupov and P Walter and R Marquet and C Ehresmann and B Ehresmann and P Dumas},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10089425},
isbn = {10089425},
year = {1999},
date = {1999-01-01},
journal = {Acta Crystallogr D Biol Crystallogr},
volume = {55},
number = {Pt 1},
pages = {281-284},
abstract = {The genomic RNA of all retroviruses is encapsidated in virions as a dimer of single-stranded chains held together near their 5'-end. For HIV-1, the initial site of dimerization has been shown to be a hairpin with a nine-residue loop containing a self-complementary sequence of six residues. This structure is proposed to promote dimerization by loop-loop interaction and formation of a so-called 'kissing complex'. A 23-nucleotide RNA strand containing the loop enclosed by a seven base-pair stem has been synthesized. This oligomer was crystallized by the vapour-diffusion method at 310 K, pH 6.5, with methyl-pentanediol as the precipitant agent in the presence of MgCl2, KCl and spermine. Quasi-complete diffraction data were obtained at 2.7 A resolution with a conventional X-ray source and at 2.3 A resolution on a synchrotron beamline. The space group is P3121 or its enantiomorph P3221, with cell parameters a = b = 60. 1},
note = {0907-4449
Journal Article},
keywords = {Base Sequence Binding Sites Crystallization Crystallography, MARQUET, Non-U.S. Gov't, Unité ARN, Viral HIV-1/*chemistry/genetics Human Molecular Sequence Data Nucleic Acid Conformation RNA, Viral/*chemistry/genetics/isolation & purification Solutions Support, X-Ray Dimerization Genome},
pubstate = {published},
tppubtype = {article}
}
The genomic RNA of all retroviruses is encapsidated in virions as a dimer of single-stranded chains held together near their 5'-end. For HIV-1, the initial site of dimerization has been shown to be a hairpin with a nine-residue loop containing a self-complementary sequence of six residues. This structure is proposed to promote dimerization by loop-loop interaction and formation of a so-called 'kissing complex'. A 23-nucleotide RNA strand containing the loop enclosed by a seven base-pair stem has been synthesized. This oligomer was crystallized by the vapour-diffusion method at 310 K, pH 6.5, with methyl-pentanediol as the precipitant agent in the presence of MgCl2, KCl and spermine. Quasi-complete diffraction data were obtained at 2.7 A resolution with a conventional X-ray source and at 2.3 A resolution on a synchrotron beamline. The space group is P3121 or its enantiomorph P3221, with cell parameters a = b = 60. 1
Wolfson A D, Khvorova A M, Sauter C, Florentz C, Giege R
Mimics of yeast tRNAAsp and their recognition by aspartyl-tRNA synthetase Article de journal
Dans: Biochemistry, vol. 38, no. 37, p. 11926-11932, 1999, ISBN: 10508395, (0006-2960 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Acylation Aspartate-tRNA Ligase/*chemistry/metabolism Base Sequence Catalysis Cloning, Asp/*chemistry/genetics/metabolism Saccharomyces cerevisiae Support, FLORENTZ, Molecular Enzyme Activation/genetics Genetic Engineering Molecular Mimicry Molecular Sequence Data Mutagenesis, Non-U.S. Gov't, Site-Directed Plasmids/chemical synthesis RNA, Transfer, Unité ARN
@article{,
title = {Mimics of yeast tRNAAsp and their recognition by aspartyl-tRNA synthetase},
author = {A D Wolfson and A M Khvorova and C Sauter and C Florentz and R Giege},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10508395},
isbn = {10508395},
year = {1999},
date = {1999-01-01},
journal = {Biochemistry},
volume = {38},
number = {37},
pages = {11926-11932},
abstract = {Assuming that the L-shaped three-dimensional structure of tRNA is an architectural framework allowing the proper presentation of identity nucleotides to aminoacyl-tRNA synthetases implies that altered and/or simplified RNA architectures can fulfill this role and be functional substrates of these enzymes, provided they contain correctly located identity elements. In this work, this paradigm was submitted to new experimental verification. Yeast aspartyl-tRNA synthetase was the model synthetase, and the extent to which the canonical structural framework of cognate tRNAAsp can be altered without losing its ability to be aminoacylated was investigated. Three novel architectures recognized by the synthetase were found. The first resembles that of metazoan mitochondrial tRNASer lacking the D-arm. The second lacks both the D- and T-arms, and the 5'-strand of the amino acid acceptor arm. The third structure is a construct in which the acceptor and anticodon helices are joined by two connectors. Aspartylation specificity of these RNAs is verified by the loss of aminoacylation activity upon mutation of the putative identity residues. Kinetic data indicate that the first two architectures are mimics of the whole tRNAAsp molecule, while the third one behaves as an aspartate minihelix mimic. Results confirm the primordial role of the discriminator nucleotide G73 in aspartylation and demonstrate that neither a helical structure in the acceptor domain nor the presence of a D- or T-arm is mandatory for specific aspartylation, but that activity relies on the presence of the cognate aspartate GUC sequence in the anticodon loop.},
note = {0006-2960
Journal Article},
keywords = {Acylation Aspartate-tRNA Ligase/*chemistry/metabolism Base Sequence Catalysis Cloning, Asp/*chemistry/genetics/metabolism Saccharomyces cerevisiae Support, FLORENTZ, Molecular Enzyme Activation/genetics Genetic Engineering Molecular Mimicry Molecular Sequence Data Mutagenesis, Non-U.S. Gov't, Site-Directed Plasmids/chemical synthesis RNA, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Assuming that the L-shaped three-dimensional structure of tRNA is an architectural framework allowing the proper presentation of identity nucleotides to aminoacyl-tRNA synthetases implies that altered and/or simplified RNA architectures can fulfill this role and be functional substrates of these enzymes, provided they contain correctly located identity elements. In this work, this paradigm was submitted to new experimental verification. Yeast aspartyl-tRNA synthetase was the model synthetase, and the extent to which the canonical structural framework of cognate tRNAAsp can be altered without losing its ability to be aminoacylated was investigated. Three novel architectures recognized by the synthetase were found. The first resembles that of metazoan mitochondrial tRNASer lacking the D-arm. The second lacks both the D- and T-arms, and the 5'-strand of the amino acid acceptor arm. The third structure is a construct in which the acceptor and anticodon helices are joined by two connectors. Aspartylation specificity of these RNAs is verified by the loss of aminoacylation activity upon mutation of the putative identity residues. Kinetic data indicate that the first two architectures are mimics of the whole tRNAAsp molecule, while the third one behaves as an aspartate minihelix mimic. Results confirm the primordial role of the discriminator nucleotide G73 in aspartylation and demonstrate that neither a helical structure in the acceptor domain nor the presence of a D- or T-arm is mandatory for specific aspartylation, but that activity relies on the presence of the cognate aspartate GUC sequence in the anticodon loop.
Wilhelm M, Heyman T, Boutabout M, Wilhelm F X
A sequence immediately upstream of the plus-strand primer is essential for plus-strand DNA synthesis of the Saccharomyces cerevisiae Ty1 retrotransposon Article de journal
Dans: Nucleic Acids Res, vol. 27, no. 23, p. 4547-4552, 1999, ISBN: 10556309, (0305-1048 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Base Sequence *DNA Primers DNA, Fungal/*biosynthesis Mutation Response Elements *Retroelements Saccharomyces cerevisiae/*genetics Support, Non-U.S. Gov't, Unité ARN
@article{,
title = {A sequence immediately upstream of the plus-strand primer is essential for plus-strand DNA synthesis of the Saccharomyces cerevisiae Ty1 retrotransposon},
author = {M Wilhelm and T Heyman and M Boutabout and F X Wilhelm},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10556309},
isbn = {10556309},
year = {1999},
date = {1999-01-01},
journal = {Nucleic Acids Res},
volume = {27},
number = {23},
pages = {4547-4552},
abstract = {Priming of plus-strand DNA is a critical step in reverse transcription of retroviruses and retrotransposons. All retroelements use an RNase H-resistant oligoribonucleotide spanning a purine-rich sequence (the polypurine tract or PPT) to prime plus-strand DNA synthesis. Plus-strand DNA synthesis of the yeast Saccharomyces cerevisiae Ty1-H3 retrotransposon is initiated at two sites, PPT1 and PPT2, located at the upstream boundary of the 3'-long terminal repeat and near the middle of the pol gene in the integrase coding region. The two plus-strand primers have the same purine-rich sequence GGGTGGTA. This sequence is not sufficient by itself to generate a plus-strand origin since two identical sequences located upstream of PPT2 in the integrase coding region are not used efficiently as primers for plus-strand DNA synthesis. Thus, other factors must be involved in the formation of a specific plus-strand DNA primer. We show here that mutations upstream of the PPT in a highly conserved T-rich region severely alters plus-strand DNA priming of Ty1. Our results demonstrate the importance of sequences or structural elements upstream of the PPT for initiation of plus-strand DNA synthesis.},
note = {0305-1048
Journal Article},
keywords = {Base Sequence *DNA Primers DNA, Fungal/*biosynthesis Mutation Response Elements *Retroelements Saccharomyces cerevisiae/*genetics Support, Non-U.S. Gov't, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Priming of plus-strand DNA is a critical step in reverse transcription of retroviruses and retrotransposons. All retroelements use an RNase H-resistant oligoribonucleotide spanning a purine-rich sequence (the polypurine tract or PPT) to prime plus-strand DNA synthesis. Plus-strand DNA synthesis of the yeast Saccharomyces cerevisiae Ty1-H3 retrotransposon is initiated at two sites, PPT1 and PPT2, located at the upstream boundary of the 3'-long terminal repeat and near the middle of the pol gene in the integrase coding region. The two plus-strand primers have the same purine-rich sequence GGGTGGTA. This sequence is not sufficient by itself to generate a plus-strand origin since two identical sequences located upstream of PPT2 in the integrase coding region are not used efficiently as primers for plus-strand DNA synthesis. Thus, other factors must be involved in the formation of a specific plus-strand DNA primer. We show here that mutations upstream of the PPT in a highly conserved T-rich region severely alters plus-strand DNA priming of Ty1. Our results demonstrate the importance of sequences or structural elements upstream of the PPT for initiation of plus-strand DNA synthesis.
Walter F, Vicens Q, Westhof E
Aminoglycoside-RNA interactions Article de journal
Dans: Curr Opin Chem Biol, vol. 3, no. 6, p. 694-704, 1999, ISBN: 10600721, (1367-5931 Journal Article Review Review, Tutorial).
Résumé | Liens | BibTeX | Étiquettes: Aminoglycosides Anti-Bacterial Agents/metabolism/*pharmacology Carbohydrate Sequence Molecular Sequence Data RNA/chemistry/*drug effects/metabolism RNA, Catalytic/antagonists & inhibitors/drug effects Support, Non-U.S. Gov't, Unité ARN
@article{,
title = {Aminoglycoside-RNA interactions},
author = {F Walter and Q Vicens and E Westhof},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10600721},
isbn = {10600721},
year = {1999},
date = {1999-01-01},
journal = {Curr Opin Chem Biol},
volume = {3},
number = {6},
pages = {694-704},
abstract = {The structural and physico-chemical parameters promoting the binding of aminoglycosides to RNAs are becoming clear. The strength of the interaction is dominated by electrostatics, with the positively charged aminoglycosides displacing metal ions. Although aminoglycosides inhibit most known ribozymes, aminoglycosides or polyamines are able to catalyze specific RNA cleavage in the absence of metal ions.},
note = {1367-5931
Journal Article
Review
Review, Tutorial},
keywords = {Aminoglycosides Anti-Bacterial Agents/metabolism/*pharmacology Carbohydrate Sequence Molecular Sequence Data RNA/chemistry/*drug effects/metabolism RNA, Catalytic/antagonists & inhibitors/drug effects Support, Non-U.S. Gov't, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
The structural and physico-chemical parameters promoting the binding of aminoglycosides to RNAs are becoming clear. The strength of the interaction is dominated by electrostatics, with the positively charged aminoglycosides displacing metal ions. Although aminoglycosides inhibit most known ribozymes, aminoglycosides or polyamines are able to catalyze specific RNA cleavage in the absence of metal ions.
Sauter C, Lorber B, Kern D, Cavarelli J, Moras D, Giege R
Crystallogenesis studies on yeast aspartyl-tRNA synthetase: use of phase diagram to improve crystal quality Article de journal
Dans: Acta Crystallogr D Biol Crystallogr, vol. 55, no. Pt 1, p. 149-156, 1999, (0907-4449 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Aspartate-tRNA Ligase/*chemistry/genetics/*isolation & purification Crystallization Crystallography, Fungal Saccharomyces cerevisiae/*enzymology/genetics Sequence Deletion Solutions Support, Non-U.S. Gov't, Unité ARN, X-Ray Genes
@article{,
title = {Crystallogenesis studies on yeast aspartyl-tRNA synthetase: use of phase diagram to improve crystal quality},
author = {C Sauter and B Lorber and D Kern and J Cavarelli and D Moras and R Giege},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10089405},
doi = {10089405},
year = {1999},
date = {1999-01-01},
journal = {Acta Crystallogr D Biol Crystallogr},
volume = {55},
number = {Pt 1},
pages = {149-156},
abstract = {Aspartyl-tRNA synthetase (AspRS) extracted from yeast is heterogeneous owing to proteolysis of its positively charged N-terminus; its crystals are of poor quality. To overcome this drawback, a rational strategy was developed to grow crystals of sufficient quality for structure determination. The strategy is based on improvement of the protein homogeneity and optimization of crystallization, taking advantage of predictions from crystal-growth theories. An active mutant lacking the first 70 residues was produced and initial crystallization conditions searched. The shape and habit of initial crystals were improved by establishing a phase diagram of protein versus crystallizing-agent concentrations. Growth of large well faceted crystals takes place at low supersaturations near the isochronic supersolubility curve. Further refinement led to reproducible growth of two crystalline forms of bipyramidal (I) or prismatic (II) habit. Both diffract X-rays better than crystals previously obtained with native AspRS. Complete data sets were collected at 3 A resolution for form I (space group P41212) and form II (space group P3221) and molecular-replacement solutions were found in both space groups.},
note = {0907-4449
Journal Article},
keywords = {Aspartate-tRNA Ligase/*chemistry/genetics/*isolation & purification Crystallization Crystallography, Fungal Saccharomyces cerevisiae/*enzymology/genetics Sequence Deletion Solutions Support, Non-U.S. Gov't, Unité ARN, X-Ray Genes},
pubstate = {published},
tppubtype = {article}
}
Aspartyl-tRNA synthetase (AspRS) extracted from yeast is heterogeneous owing to proteolysis of its positively charged N-terminus; its crystals are of poor quality. To overcome this drawback, a rational strategy was developed to grow crystals of sufficient quality for structure determination. The strategy is based on improvement of the protein homogeneity and optimization of crystallization, taking advantage of predictions from crystal-growth theories. An active mutant lacking the first 70 residues was produced and initial crystallization conditions searched. The shape and habit of initial crystals were improved by establishing a phase diagram of protein versus crystallizing-agent concentrations. Growth of large well faceted crystals takes place at low supersaturations near the isochronic supersolubility curve. Further refinement led to reproducible growth of two crystalline forms of bipyramidal (I) or prismatic (II) habit. Both diffract X-rays better than crystals previously obtained with native AspRS. Complete data sets were collected at 3 A resolution for form I (space group P41212) and form II (space group P3221) and molecular-replacement solutions were found in both space groups.
Rudinger-Thirion J, Giege R
The peculiar architectural framework of tRNASec is fully recognized by yeast AspRS Article de journal
Dans: RNA, vol. 5, no. 4, p. 495-502, 1999, ISBN: 10199566, (1355-8382 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Amino Acid-Specific/*genetics RNA, Amino Acyl-tRNA Ligases/*genetics/metabolism Anticodon/genetics Aspartic Acid/genetics/metabolism Base Sequence Escherichia coli/genetics Fungi/*enzymology/genetics Molecular Sequence Data Mutation Nucleic Acid Conformation RNA, Asp/genetics Selenocysteine/genetics/metabolism Support, Bacterial/genetics RNA, Non-U.S. Gov't, Transfer, Unité ARN
@article{,
title = {The peculiar architectural framework of tRNASec is fully recognized by yeast AspRS},
author = {J Rudinger-Thirion and R Giege},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10199566},
isbn = {10199566},
year = {1999},
date = {1999-01-01},
journal = {RNA},
volume = {5},
number = {4},
pages = {495-502},
abstract = {The wild-type transcript of Escherichia coli tRNASec, characterized by a peculiar core architecture and a large variable region, was shown to be aspartylatable by yeast AspRS. Similar activities were found for tRNASec mutants with methionine, leucine, and tryptophan anticodons. The charging efficiency of these molecules was found comparable to that of a minihelix derived from tRNAAsp and is accounted for by the presence of the discriminator residue G73, which is a major aspartate identity determinant. Introducing the aspartate identity elements from the anticodon loop (G34, U35, C36, C38) into tRNASec transforms this molecule into an aspartate acceptor with kinetic properties identical to tRNAAsp. Expression of the aspartate identity set in tRNASec is independent of the size of its variable region. The functional study was completed by footprinting experiments with four different nucleases as structural probes. Protection patterns by AspRS of transplanted tRNASec and tRNAAsp were found similar. They are modified, particularly in the anticodon loop, upon changing the aspartate anticodon into that of methionine. Altogether, it appears that recognition of a tRNA by AspRS is more governed by the presence of the aspartate identity set than by the structural framework that carries this set.},
note = {1355-8382
Journal Article},
keywords = {Amino Acid-Specific/*genetics RNA, Amino Acyl-tRNA Ligases/*genetics/metabolism Anticodon/genetics Aspartic Acid/genetics/metabolism Base Sequence Escherichia coli/genetics Fungi/*enzymology/genetics Molecular Sequence Data Mutation Nucleic Acid Conformation RNA, Asp/genetics Selenocysteine/genetics/metabolism Support, Bacterial/genetics RNA, Non-U.S. Gov't, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
The wild-type transcript of Escherichia coli tRNASec, characterized by a peculiar core architecture and a large variable region, was shown to be aspartylatable by yeast AspRS. Similar activities were found for tRNASec mutants with methionine, leucine, and tryptophan anticodons. The charging efficiency of these molecules was found comparable to that of a minihelix derived from tRNAAsp and is accounted for by the presence of the discriminator residue G73, which is a major aspartate identity determinant. Introducing the aspartate identity elements from the anticodon loop (G34, U35, C36, C38) into tRNASec transforms this molecule into an aspartate acceptor with kinetic properties identical to tRNAAsp. Expression of the aspartate identity set in tRNASec is independent of the size of its variable region. The functional study was completed by footprinting experiments with four different nucleases as structural probes. Protection patterns by AspRS of transplanted tRNASec and tRNAAsp were found similar. They are modified, particularly in the anticodon loop, upon changing the aspartate anticodon into that of methionine. Altogether, it appears that recognition of a tRNA by AspRS is more governed by the presence of the aspartate identity set than by the structural framework that carries this set.
Petyuk V A, Zenkova M A, Giege R, Vlassov V V
Hybridization of antisense oligonucleotides with the 3'part of tRNA(Phe) Article de journal
Dans: FEBS Lett, vol. 444, no. 2-3, p. 217-221, 1999, ISBN: 10050762, (0014-5793 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Antisense/*genetics RNA, Base Sequence Electrophoresis, Calf Thymus/metabolism Support, Fungal/genetics RNA, Non-U.S. Gov't, Phe/*genetics Ribonuclease H, Polyacrylamide Gel Molecular Sequence Data Nucleic Acid Conformation Nucleic Acid Hybridization/*genetics Oligodeoxyribonucleotides/genetics Oligonucleotides, Transfer, Unité ARN
@article{,
title = {Hybridization of antisense oligonucleotides with the 3'part of tRNA(Phe)},
author = {V A Petyuk and M A Zenkova and R Giege and V V Vlassov},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10050762},
isbn = {10050762},
year = {1999},
date = {1999-01-01},
journal = {FEBS Lett},
volume = {444},
number = {2-3},
pages = {217-221},
abstract = {The interaction of antisense oligodeoxyribonucleotides with yeast tRNA(Phe) was investigated. 14-15-mers complementary to the 3'-terminal sequence including the ACCA end bind to the tRNA under physiological conditions. At low oligonucleotide concentrations the binding occurs at the unique complementary site. At higher oligonucleotide concentrations, the second oligonucleotide molecule binds to the complex due to non-perfect duplex formation in the T-loop stabilized by stacking between the two bound oligonucleotides. In these complexes the acceptor stem is open and the 5'-terminal sequence of the tRNA is accessible for binding of a complementary oligonucleotide. The results prove that the efficient binding of oligonucleotides to the 3'-terminal sequence of the tRNA occurs through initial binding to the single-stranded sequence ACCA followed by invasion in the acceptor stem and strand displacement.},
note = {0014-5793
Journal Article},
keywords = {Antisense/*genetics RNA, Base Sequence Electrophoresis, Calf Thymus/metabolism Support, Fungal/genetics RNA, Non-U.S. Gov't, Phe/*genetics Ribonuclease H, Polyacrylamide Gel Molecular Sequence Data Nucleic Acid Conformation Nucleic Acid Hybridization/*genetics Oligodeoxyribonucleotides/genetics Oligonucleotides, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
The interaction of antisense oligodeoxyribonucleotides with yeast tRNA(Phe) was investigated. 14-15-mers complementary to the 3'-terminal sequence including the ACCA end bind to the tRNA under physiological conditions. At low oligonucleotide concentrations the binding occurs at the unique complementary site. At higher oligonucleotide concentrations, the second oligonucleotide molecule binds to the complex due to non-perfect duplex formation in the T-loop stabilized by stacking between the two bound oligonucleotides. In these complexes the acceptor stem is open and the 5'-terminal sequence of the tRNA is accessible for binding of a complementary oligonucleotide. The results prove that the efficient binding of oligonucleotides to the 3'-terminal sequence of the tRNA occurs through initial binding to the single-stranded sequence ACCA followed by invasion in the acceptor stem and strand displacement.
Petyuk V, Zenkova M, Giege R, Vlassov V
Interaction of complementary oligonucleotides with the 3'-end of yeast tRNA(Phe) Article de journal
Dans: Nucleosides Nucleotides, vol. 18, no. 6-7, p. 1459-1461, 1999, ISBN: 10474225, (0732-8311 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Non-U.S. Gov't, Nucleic Acid Conformation RNA, Phe/*chemistry Saccharomyces cerevisiae/*genetics Support, Transfer, Unité ARN
@article{,
title = {Interaction of complementary oligonucleotides with the 3'-end of yeast tRNA(Phe)},
author = {V Petyuk and M Zenkova and R Giege and V Vlassov},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10474225},
isbn = {10474225},
year = {1999},
date = {1999-01-01},
journal = {Nucleosides Nucleotides},
volume = {18},
number = {6-7},
pages = {1459-1461},
abstract = {Interaction of yeast tRNA(Phe) with oligodeoxyribonucleotides (ONs), complementary to the nucleotides 62-76 was investigated. Results of gel-mobility shift assay and RNase A probing evidence that the ONs containing the sequence complementary to the tRNA ACCA end can easily invade the hairpin structure under physiological conditions. The limiting step of association process is the tRNA unfolding.},
note = {0732-8311
Journal Article},
keywords = {Non-U.S. Gov't, Nucleic Acid Conformation RNA, Phe/*chemistry Saccharomyces cerevisiae/*genetics Support, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Interaction of yeast tRNA(Phe) with oligodeoxyribonucleotides (ONs), complementary to the nucleotides 62-76 was investigated. Results of gel-mobility shift assay and RNase A probing evidence that the ONs containing the sequence complementary to the tRNA ACCA end can easily invade the hairpin structure under physiological conditions. The limiting step of association process is the tRNA unfolding.
Perrin L, Romby P, Laurenti P, Berenger H, Kallenbach S, Bourbon H M, Pradel J
Dans: J Biol Chem, vol. 274, no. 10, p. 6315-6323, 1999, ISBN: 10037720, (0021-9258 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Animals Base Sequence Binding Sites/genetics Chromatin/*genetics DNA/*genetics DNA-Binding Proteins/*genetics/metabolism Drosophila/*genetics *Drosophila Proteins *Genes, Insect Insect Proteins/*genetics/metabolism Molecular Sequence Data Phosphorylation RNA/*genetics RNA-Binding Proteins/*genetics/metabolism Support, Non-U.S. Gov't, ROMBY, Unité ARN
@article{,
title = {The Drosophila modifier of variegation modulo gene product binds specific RNA sequences at the nucleolus and interacts with DNA and chromatin in a phosphorylation-dependent manner},
author = {L Perrin and P Romby and P Laurenti and H Berenger and S Kallenbach and H M Bourbon and J Pradel},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10037720},
isbn = {10037720},
year = {1999},
date = {1999-01-01},
journal = {J Biol Chem},
volume = {274},
number = {10},
pages = {6315-6323},
abstract = {modulo belongs to the modifier of Position Effect Variegation class of Drosophila genes, suggesting a role for its product in regulating chromatin structure. Genetics assigned a second function to the gene, in protein synthesis capacity. Bifunctionality is consistent with protein localization in two distinct subnuclear compartments, chromatin and nucleolus, and with its organization in modules potentially involved in DNA and RNA binding. In this study, we examine nucleic acid interactions established by Modulo at nucleolus and chromatin and the mechanism that controls the distribution and balances the function of the protein in the two compartments. Structure/function analysis and oligomer selection/amplification experiments indicate that, in vitro, two basic terminal domains independently contact DNA without sequence specificity, whereas a central RNA Recognition Motif (RRM)-containing domain allows recognition of a novel sequence-/motif-specific RNA class. Phosphorylation moreover is shown to down-regulate DNA binding. Evidence is provided that in vivo nucleolar Modulo is highly phosphorylated and belongs to a ribonucleoprotein particle, whereas chromatin-associated protein is not modified. A functional scheme is finally proposed in which modification by phosphorylation modulates Mod subnuclear distribution and balances its function at the nucleolus and chromatin.},
note = {0021-9258
Journal Article},
keywords = {Animals Base Sequence Binding Sites/genetics Chromatin/*genetics DNA/*genetics DNA-Binding Proteins/*genetics/metabolism Drosophila/*genetics *Drosophila Proteins *Genes, Insect Insect Proteins/*genetics/metabolism Molecular Sequence Data Phosphorylation RNA/*genetics RNA-Binding Proteins/*genetics/metabolism Support, Non-U.S. Gov't, ROMBY, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
modulo belongs to the modifier of Position Effect Variegation class of Drosophila genes, suggesting a role for its product in regulating chromatin structure. Genetics assigned a second function to the gene, in protein synthesis capacity. Bifunctionality is consistent with protein localization in two distinct subnuclear compartments, chromatin and nucleolus, and with its organization in modules potentially involved in DNA and RNA binding. In this study, we examine nucleic acid interactions established by Modulo at nucleolus and chromatin and the mechanism that controls the distribution and balances the function of the protein in the two compartments. Structure/function analysis and oligomer selection/amplification experiments indicate that, in vitro, two basic terminal domains independently contact DNA without sequence specificity, whereas a central RNA Recognition Motif (RRM)-containing domain allows recognition of a novel sequence-/motif-specific RNA class. Phosphorylation moreover is shown to down-regulate DNA binding. Evidence is provided that in vivo nucleolar Modulo is highly phosphorylated and belongs to a ribonucleoprotein particle, whereas chromatin-associated protein is not modified. A functional scheme is finally proposed in which modification by phosphorylation modulates Mod subnuclear distribution and balances its function at the nucleolus and chromatin.
Ennifar E, Yusupov M, Walter P, Marquet R, Ehresmann B, Ehresmann C, Dumas P
The crystal structure of the dimerization initiation site of genomic HIV-1 RNA reveals an extended duplex with two adenine bulges Article de journal
Dans: Structure, vol. 7, no. 11, p. 1439-49, 1999, ISBN: 10574792, (0969-2126 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Adenine/*chemistry Base Pair Mismatch Base Sequence Crystallography, ENNIFAR, Molecular Molecular Sequence Data *Nucleic Acid Conformation RNA, Non-U.S. Gov't, Unité ARN, Viral/*chemistry/metabolism Support, X-Ray Dimerization HIV-1/*genetics Magnesium/metabolism Magnetic Resonance Spectroscopy Manganese/metabolism Models
@article{,
title = {The crystal structure of the dimerization initiation site of genomic HIV-1 RNA reveals an extended duplex with two adenine bulges},
author = {E Ennifar and M Yusupov and P Walter and R Marquet and B Ehresmann and C Ehresmann and P Dumas},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10574792},
isbn = {10574792},
year = {1999},
date = {1999-01-01},
journal = {Structure},
volume = {7},
number = {11},
pages = {1439-49},
abstract = {BACKGROUND: An important step in retroviral replication is dimerization of the genomic RNA prior to encapsidation. Dimerization is initiated by the formation of a transient 'kissing-loop complex' that is thought to be subsequently matured into an extended duplex by the nucleocapsid protein (NCp). Although chemical probing and nuclear magnetic resonance spectroscopy have provided insight into the structure of the kissing-loop structure, no structural information concerning the extended-duplex state is available so far. RESULTS: The structure of a minimal HIV-1 RNA dimerization initiation site has been solved at 2.3 A resolution in two different space groups. It reveals a 22 base pair extended duplex with two noncanonical Watson-Crick-like G-A mismatches, each adjacent to a bulged-out adenine. The structure shows significant asymmetry in deep groove width and G-A base-pair conformations. A network of eight magnesium cations was clearly identified, one being unusually chelated by the 3' phosphate of each bulge across an extremely narrowed deep major groove. CONCLUSIONS: These crystal structures represent the putative matured form of the initial kissing-loop complex. They show the ability of this self-complementary RNA hairpin loop to acquire a more stable extended duplex structure. Both bulged adenines form a striking 'base grip' that could be a recognition signal, either in cis for another viral RNA sequence, or in trans for a protein, possibly the NCp. Magnesium binding might be important to promote and stabilize the observed extrahelical conformation of these bulges.},
note = {0969-2126
Journal Article},
keywords = {Adenine/*chemistry Base Pair Mismatch Base Sequence Crystallography, ENNIFAR, Molecular Molecular Sequence Data *Nucleic Acid Conformation RNA, Non-U.S. Gov't, Unité ARN, Viral/*chemistry/metabolism Support, X-Ray Dimerization HIV-1/*genetics Magnesium/metabolism Magnetic Resonance Spectroscopy Manganese/metabolism Models},
pubstate = {published},
tppubtype = {article}
}
BACKGROUND: An important step in retroviral replication is dimerization of the genomic RNA prior to encapsidation. Dimerization is initiated by the formation of a transient 'kissing-loop complex' that is thought to be subsequently matured into an extended duplex by the nucleocapsid protein (NCp). Although chemical probing and nuclear magnetic resonance spectroscopy have provided insight into the structure of the kissing-loop structure, no structural information concerning the extended-duplex state is available so far. RESULTS: The structure of a minimal HIV-1 RNA dimerization initiation site has been solved at 2.3 A resolution in two different space groups. It reveals a 22 base pair extended duplex with two noncanonical Watson-Crick-like G-A mismatches, each adjacent to a bulged-out adenine. The structure shows significant asymmetry in deep groove width and G-A base-pair conformations. A network of eight magnesium cations was clearly identified, one being unusually chelated by the 3' phosphate of each bulge across an extremely narrowed deep major groove. CONCLUSIONS: These crystal structures represent the putative matured form of the initial kissing-loop complex. They show the ability of this self-complementary RNA hairpin loop to acquire a more stable extended duplex structure. Both bulged adenines form a striking 'base grip' that could be a recognition signal, either in cis for another viral RNA sequence, or in trans for a protein, possibly the NCp. Magnesium binding might be important to promote and stabilize the observed extrahelical conformation of these bulges.
1997
Walczak R, Hubert N, Carbon P, Krol A
Solution structure of SECIS, the mRNA element required for eukaryotic selenocysteine insertion--interaction studies with the SECIS-binding protein SBP Article de journal
Dans: Biomed Environ Sci, vol. 10, no. 2-3, p. 177-181, 1997, ISBN: 9315308, (0895-3988 Journal Article Review Review, Tutorial).
Résumé | Liens | BibTeX | Étiquettes: Animals Human Magnetic Resonance Spectroscopy Models, Chemical Nucleic Acid Conformation RNA, Messenger/*chemistry Rats Selenocysteine/*chemistry Solutions Support, Non-U.S. Gov't, Unité ARN
@article{,
title = {Solution structure of SECIS, the mRNA element required for eukaryotic selenocysteine insertion--interaction studies with the SECIS-binding protein SBP},
author = {R Walczak and N Hubert and P Carbon and A Krol},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9315308},
isbn = {9315308},
year = {1997},
date = {1997-01-01},
journal = {Biomed Environ Sci},
volume = {10},
number = {2-3},
pages = {177-181},
abstract = {Selenocysteine, a selenium-containing analog of cysteine, is found in the prokaryotic and eukaryotic kingdoms in active sites of enzymes involved in oxidation-reduction reactions. This aminoacid is cotranslationally incorporated at UGA codons which usually act as translation stop codons. In eukaryotes, decoding of selenocysteine necessitates the participation of the selenocysteine insertion sequence (SECIS), an element lying in the 3'-untranslated region of selenoprotein mRNAs. A detailed experimental study of the secondary structures of the SECIS elements of rat and human type 1 iodothyronine deiodinases and rat glutathione peroxidase was performed. Enzymatic and chemical structure probing led us to propose a secondary structure model, supported by sequence comparison of 23 SECIS mRNAs. The secondary structure model revealed the existence of a novel type of RNA motif composed of four consecutive non-Watson-Crick base-pairs. Using gel shift experiments, we identified in several mammalian cell type extracts the protein SBP, for SECIS-binding protein, that specifically recognizes the iodothyronine deiodinases and glutathione peroxidase SECIS elements. The structural model that we derived for the SECIS RNAs discloses RNA features possibly implicated in the binding of SBP and/or SECIS function.},
note = {0895-3988
Journal Article
Review
Review, Tutorial},
keywords = {Animals Human Magnetic Resonance Spectroscopy Models, Chemical Nucleic Acid Conformation RNA, Messenger/*chemistry Rats Selenocysteine/*chemistry Solutions Support, Non-U.S. Gov't, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Selenocysteine, a selenium-containing analog of cysteine, is found in the prokaryotic and eukaryotic kingdoms in active sites of enzymes involved in oxidation-reduction reactions. This aminoacid is cotranslationally incorporated at UGA codons which usually act as translation stop codons. In eukaryotes, decoding of selenocysteine necessitates the participation of the selenocysteine insertion sequence (SECIS), an element lying in the 3'-untranslated region of selenoprotein mRNAs. A detailed experimental study of the secondary structures of the SECIS elements of rat and human type 1 iodothyronine deiodinases and rat glutathione peroxidase was performed. Enzymatic and chemical structure probing led us to propose a secondary structure model, supported by sequence comparison of 23 SECIS mRNAs. The secondary structure model revealed the existence of a novel type of RNA motif composed of four consecutive non-Watson-Crick base-pairs. Using gel shift experiments, we identified in several mammalian cell type extracts the protein SBP, for SECIS-binding protein, that specifically recognizes the iodothyronine deiodinases and glutathione peroxidase SECIS elements. The structural model that we derived for the SECIS RNAs discloses RNA features possibly implicated in the binding of SBP and/or SECIS function.
Sissler M, Eriani G, Martin F, Giege R, Florentz C
Mirror image alternative interaction patterns of the same tRNA with either class I arginyl-tRNA synthetase or class II aspartyl-tRNA synthetase Article de journal
Dans: Nucleic Acids Res, vol. 25, no. 24, p. 4899-4906, 1997, ISBN: 9396794, (0305-1048 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Anticodon/chemistry Arginine-tRNA Ligase/classification/*metabolism Aspartate-tRNA Ligase/classification/*metabolism Base Sequence DNA Footprinting Escherichia coli Fungal Proteins/classification/*metabolism Models, Arg/chemistry/*metabolism RNA, Asp/chemistry/*metabolism Recombinant Fusion Proteins/metabolism Saccharomyces cerevisiae/metabolism Stereoisomerism Substrate Specificity Support, ERIANI, FLORENTZ, Fungal/chemistry/*metabolism RNA, Molecular Molecular Sequence Data *Nucleic Acid Conformation Protein Binding RNA, Non-U.S. Gov't, SISSLER, Transfer, Unité ARN
@article{,
title = {Mirror image alternative interaction patterns of the same tRNA with either class I arginyl-tRNA synthetase or class II aspartyl-tRNA synthetase},
author = {M Sissler and G Eriani and F Martin and R Giege and C Florentz},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9396794},
isbn = {9396794},
year = {1997},
date = {1997-01-01},
journal = {Nucleic Acids Res},
volume = {25},
number = {24},
pages = {4899-4906},
abstract = {Gene cloning, overproduction and an efficient purification protocol of yeast arginyl-tRNA synthetase (ArgRS) as well as the interaction patterns of this protein with cognate tRNAArgand non-cognate tRNAAspare described. This work was motivated by the fact that the in vitro transcript of tRNAAspis of dual aminoacylation specificity and is not only aspartylated but also efficiently arginylated. The crystal structure of the complex between class II aspartyl-tRNA synthetase (AspRS) and tRNAAsp, as well as early biochemical data, have shown that tRNAAspis recognized by its variable region side. Here we show by footprinting with enzymatic and chemical probes that transcribed tRNAAspis contacted by class I ArgRS along the opposite D arm side, as is homologous tRNAArg, but with idiosyncratic interaction patterns. Besides protection, footprints also show enhanced accessibility of the tRNAs to the structural probes, indicative of conformational changes in the complexed tRNAs. These different patterns are interpreted in relation to the alternative arginine identity sets found in the anticodon loops of tRNAArgand tRNAAsp. The mirror image alternative interaction patterns of unmodified tRNAAspwith either class I ArgRS or class II AspRS, accounting for the dual identity of this tRNA, are discussed in relation to the class defining features of the synthetases. This study indicates that complex formation between unmodified tRNAAspand either ArgRS and AspRS is solely governed by the proteins.},
note = {0305-1048
Journal Article},
keywords = {Anticodon/chemistry Arginine-tRNA Ligase/classification/*metabolism Aspartate-tRNA Ligase/classification/*metabolism Base Sequence DNA Footprinting Escherichia coli Fungal Proteins/classification/*metabolism Models, Arg/chemistry/*metabolism RNA, Asp/chemistry/*metabolism Recombinant Fusion Proteins/metabolism Saccharomyces cerevisiae/metabolism Stereoisomerism Substrate Specificity Support, ERIANI, FLORENTZ, Fungal/chemistry/*metabolism RNA, Molecular Molecular Sequence Data *Nucleic Acid Conformation Protein Binding RNA, Non-U.S. Gov't, SISSLER, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Gene cloning, overproduction and an efficient purification protocol of yeast arginyl-tRNA synthetase (ArgRS) as well as the interaction patterns of this protein with cognate tRNAArgand non-cognate tRNAAspare described. This work was motivated by the fact that the in vitro transcript of tRNAAspis of dual aminoacylation specificity and is not only aspartylated but also efficiently arginylated. The crystal structure of the complex between class II aspartyl-tRNA synthetase (AspRS) and tRNAAsp, as well as early biochemical data, have shown that tRNAAspis recognized by its variable region side. Here we show by footprinting with enzymatic and chemical probes that transcribed tRNAAspis contacted by class I ArgRS along the opposite D arm side, as is homologous tRNAArg, but with idiosyncratic interaction patterns. Besides protection, footprints also show enhanced accessibility of the tRNAs to the structural probes, indicative of conformational changes in the complexed tRNAs. These different patterns are interpreted in relation to the alternative arginine identity sets found in the anticodon loops of tRNAArgand tRNAAsp. The mirror image alternative interaction patterns of unmodified tRNAAspwith either class I ArgRS or class II AspRS, accounting for the dual identity of this tRNA, are discussed in relation to the class defining features of the synthetases. This study indicates that complex formation between unmodified tRNAAspand either ArgRS and AspRS is solely governed by the proteins.
Senger B, Auxilien S, Englisch U, Cramer F, Fasiolo F
The modified wobble base inosine in yeast tRNAIle is a positive determinant for aminoacylation by isoleucyl-tRNA synthetase Article de journal
Dans: Biochemistry, vol. 36, no. 27, p. 8269-8275, 1997, ISBN: 9204872, (0006-2960 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Acylation Anticodon Base Sequence Escherichia coli/genetics Inosine/*chemistry/metabolism Isoleucine-tRNA Ligase/*metabolism Molecular Sequence Data Nucleic Acid Conformation Pseudouridine/chemistry/metabolism RNA, Bacterial/chemistry/metabolism RNA, Fungal/chemistry/metabolism RNA, Ile/*chemistry/metabolism Saccharomyces cerevisiae/*genetics Structure-Activity Relationship Substrate Specificity Support, Non-U.S. Gov't, Transfer, Unité ARN
@article{,
title = {The modified wobble base inosine in yeast tRNAIle is a positive determinant for aminoacylation by isoleucyl-tRNA synthetase},
author = {B Senger and S Auxilien and U Englisch and F Cramer and F Fasiolo},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9204872},
isbn = {9204872},
year = {1997},
date = {1997-01-01},
journal = {Biochemistry},
volume = {36},
number = {27},
pages = {8269-8275},
abstract = {Earlier work by two independent groups has established the fact that anticodons GAU and LAU of Escherichia coli tRNAIle isoacceptors play a critical role in the tRNA identity. Yeast possesses two isoleucine transfer RNAs, a major one with anticodon IAU and a minor one with anticodon PsiAPsi which are derived from the post-transcriptional modification of AAU and UAU gene sequences, respectively. We present direct evidence which reveals that inosine is a positive determinant for yeast isoleucyl-tRNA synthetase. We also show that yeast tRNAMet with guanosine at the wobble position becomes aminoacylated with isoleucine while methionine acceptance is lost. As inosine and guanosine share the 6-keto and the N-1 hydrogen groups, this suggests that these hydrogen donor and acceptor groups are determinants for isoleucine specificity. The role of the minor tRNAIle anticodon pseudouridines in tRNA isoleucylation could not be tested directly but was deduced from a 40-fold decrease in the activity of the unmodified transcript. The presence of the NHCO structure in guanosine, inosine, pseudouridine, and lysidine suggests a unifying model of wobble base recognition by the yeast and E. coli isoleucyl-tRNA synthetase. In contrast to lysidine which switches the identity of the tRNA from methionine to isoleucine [Muramatsu, T., Nishikawa, K., Nemoto, F., Kuchino, Y., Nishimura, S., Miyazawa, T., & Yokoyama, S. (1988) Nature 336, 179-181], pseudouridine-34 does not modify the specificity of the yeast minor tRNAIle since U-34 is a strong negative determinant for yeast MetRS. Therefore, the major role of Psi-34 (in combination with Psi-36 or not) is likely in isoleucine AUA codon specificity and translational fidelity.},
note = {0006-2960
Journal Article},
keywords = {Acylation Anticodon Base Sequence Escherichia coli/genetics Inosine/*chemistry/metabolism Isoleucine-tRNA Ligase/*metabolism Molecular Sequence Data Nucleic Acid Conformation Pseudouridine/chemistry/metabolism RNA, Bacterial/chemistry/metabolism RNA, Fungal/chemistry/metabolism RNA, Ile/*chemistry/metabolism Saccharomyces cerevisiae/*genetics Structure-Activity Relationship Substrate Specificity Support, Non-U.S. Gov't, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Earlier work by two independent groups has established the fact that anticodons GAU and LAU of Escherichia coli tRNAIle isoacceptors play a critical role in the tRNA identity. Yeast possesses two isoleucine transfer RNAs, a major one with anticodon IAU and a minor one with anticodon PsiAPsi which are derived from the post-transcriptional modification of AAU and UAU gene sequences, respectively. We present direct evidence which reveals that inosine is a positive determinant for yeast isoleucyl-tRNA synthetase. We also show that yeast tRNAMet with guanosine at the wobble position becomes aminoacylated with isoleucine while methionine acceptance is lost. As inosine and guanosine share the 6-keto and the N-1 hydrogen groups, this suggests that these hydrogen donor and acceptor groups are determinants for isoleucine specificity. The role of the minor tRNAIle anticodon pseudouridines in tRNA isoleucylation could not be tested directly but was deduced from a 40-fold decrease in the activity of the unmodified transcript. The presence of the NHCO structure in guanosine, inosine, pseudouridine, and lysidine suggests a unifying model of wobble base recognition by the yeast and E. coli isoleucyl-tRNA synthetase. In contrast to lysidine which switches the identity of the tRNA from methionine to isoleucine [Muramatsu, T., Nishikawa, K., Nemoto, F., Kuchino, Y., Nishimura, S., Miyazawa, T., & Yokoyama, S. (1988) Nature 336, 179-181], pseudouridine-34 does not modify the specificity of the yeast minor tRNAIle since U-34 is a strong negative determinant for yeast MetRS. Therefore, the major role of Psi-34 (in combination with Psi-36 or not) is likely in isoleucine AUA codon specificity and translational fidelity.
Schlegl J, Gegout V, Schlager B, Hentze M W, Westhof E, Ehresmann C, Ehresmann B, Romby P
Probing the structure of the regulatory region of human transferrin receptor messenger RNA and its interaction with iron regulatory protein-1 Article de journal
Dans: RNA, vol. 3, no. 10, p. 1159-1172, 1997, ISBN: 9326491, (1355-8382 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Base Composition Base Sequence Binding Sites Electrophoresis, Messenger/*chemistry/metabolism RNA-Binding Proteins/*metabolism Receptors, Molecular Molecular Sequence Data Mutation *Nucleic Acid Conformation RNA, Non-U.S. Gov't, Polyacrylamide Gel Ethylnitrosourea/pharmacology Human Hydrolysis Hydroxyl Radical/metabolism Iron/metabolism Iron Regulatory Protein 1 Iron-Regulatory Proteins Iron-Sulfur Proteins/*metabolism Lead/pharmacology Models, ROMBY, Transferrin/*genetics Ribonuclease T1/metabolism Support, Unité ARN
@article{,
title = {Probing the structure of the regulatory region of human transferrin receptor messenger RNA and its interaction with iron regulatory protein-1},
author = {J Schlegl and V Gegout and B Schlager and M W Hentze and E Westhof and C Ehresmann and B Ehresmann and P Romby},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9326491},
isbn = {9326491},
year = {1997},
date = {1997-01-01},
journal = {RNA},
volume = {3},
number = {10},
pages = {1159-1172},
abstract = {A portion of the 3'UTR of the human transferrin receptor mRNA mediates iron-dependent regulation of mRNA stability. The minimal RNA regulatory region contains three conserved hairpins, so-called iron responsive elements (IREs), that are recognized specifically by iron regulatory proteins (IRPs). The structure of this regulatory region and its complex with IRP-1 was probed using a combination of enzymes and chemicals. The data support the existence of an intrinsic IRE loop structure that is constrained by an internal C-G base pair. This particular structure is one of the determinants required for optimal IRP binding. IRP-1 covers one helical turn of the IRE and protects conserved residues in each of the three IREs: the bulged cytosine and nucleotides in the hairpin loops. Two essential IRP-phosphate contacts were identified by ethylation interference. Three-dimensional modeling of one IRE reveals that IRP-1 contacts several bases and the ribose-phosphate backbone located on one face in the deep groove, but contacts also exist with the shallow groove. A conformational change of the IRE loop mediated by IRP-1 binding was visualized by Pb2+-catalyzed hydrolysis. This effect is dependent on the loop structure and on the nature of the closing base pair. Within the regulatory region of transferrin receptor mRNA, IRP-1 induces reactivity changes in a U-rich hairpin loop that requires the presence of the stem-loop structure located just downstream the endonucleolytic cleavage site identified by Binder et al. (Binder R et al. 1994, EMBO J 13:1969-1980). These results provide indications of the mechanism by which IRP-1 stabilizes the transferrin receptor mRNA under iron depletion conditions.},
note = {1355-8382
Journal Article},
keywords = {Base Composition Base Sequence Binding Sites Electrophoresis, Messenger/*chemistry/metabolism RNA-Binding Proteins/*metabolism Receptors, Molecular Molecular Sequence Data Mutation *Nucleic Acid Conformation RNA, Non-U.S. Gov't, Polyacrylamide Gel Ethylnitrosourea/pharmacology Human Hydrolysis Hydroxyl Radical/metabolism Iron/metabolism Iron Regulatory Protein 1 Iron-Regulatory Proteins Iron-Sulfur Proteins/*metabolism Lead/pharmacology Models, ROMBY, Transferrin/*genetics Ribonuclease T1/metabolism Support, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
A portion of the 3'UTR of the human transferrin receptor mRNA mediates iron-dependent regulation of mRNA stability. The minimal RNA regulatory region contains three conserved hairpins, so-called iron responsive elements (IREs), that are recognized specifically by iron regulatory proteins (IRPs). The structure of this regulatory region and its complex with IRP-1 was probed using a combination of enzymes and chemicals. The data support the existence of an intrinsic IRE loop structure that is constrained by an internal C-G base pair. This particular structure is one of the determinants required for optimal IRP binding. IRP-1 covers one helical turn of the IRE and protects conserved residues in each of the three IREs: the bulged cytosine and nucleotides in the hairpin loops. Two essential IRP-phosphate contacts were identified by ethylation interference. Three-dimensional modeling of one IRE reveals that IRP-1 contacts several bases and the ribose-phosphate backbone located on one face in the deep groove, but contacts also exist with the shallow groove. A conformational change of the IRE loop mediated by IRP-1 binding was visualized by Pb2+-catalyzed hydrolysis. This effect is dependent on the loop structure and on the nature of the closing base pair. Within the regulatory region of transferrin receptor mRNA, IRP-1 induces reactivity changes in a U-rich hairpin loop that requires the presence of the stem-loop structure located just downstream the endonucleolytic cleavage site identified by Binder et al. (Binder R et al. 1994, EMBO J 13:1969-1980). These results provide indications of the mechanism by which IRP-1 stabilizes the transferrin receptor mRNA under iron depletion conditions.
Rudinger J, Felden B, Florentz C, Giege R
Strategy for RNA recognition by yeast histidyl-tRNA synthetase Article de journal
Dans: Bioorg Med Chem, vol. 5, no. 6, p. 1001-1009, 1997, ISBN: 9222493, (0968-0896 Journal Article Review Review, Tutorial).
Résumé | Liens | BibTeX | Étiquettes: Amino Acyl/metabolism RNA, Base Sequence Histidine-tRNA Ligase/*metabolism Molecular Sequence Data RNA, FLORENTZ, Fungal/*metabolism RNA, Non-U.S. Gov't, Transfer, Unité ARN, Viral/metabolism Saccharomyces cerevisiae/*enzymology Substrate Specificity Support
@article{,
title = {Strategy for RNA recognition by yeast histidyl-tRNA synthetase},
author = {J Rudinger and B Felden and C Florentz and R Giege},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9222493},
isbn = {9222493},
year = {1997},
date = {1997-01-01},
journal = {Bioorg Med Chem},
volume = {5},
number = {6},
pages = {1001-1009},
abstract = {Histidine aminoacylation systems are of interest because of the structural diversity of the RNA substrates recognized by histidyl-tRNA synthetases. Among tRNAs participating in protein synthesis, those specific for histidine all share an additional residue at their 5'-extremities. On the other hand, tRNA-like domains at the 3'--termini of some plant viruses can also be charged by histidyl-tRNA synthetases, although they are not actors in protein synthesis. This is the case for the RNAs from tobacco mosaic virus and its satellite virus but also those of turnip yellow and brome mosaic viruses. All these RNAs have intricate foldings at their 3'-termini differing from that of canonical tRNAs and share a pseudoknotted domain which is the prerequisite for their folding into structures mimicking the overall L-shape of tRNAs. This paper gives an overview on tRNA identity and rationalizes the apparently contradictory structural and aminoacylation features of histidine-specific tRNAs and tRNA-like structures. The discussion mainly relies on histidylation data obtained with the yeast synthetase, but the conclusions are of a more universal nature. In canonical tRNA(His), the major histidine identity element is the 'minus' 1 residue, since its removal impairs histidylation and conversely its addition to a non-cognate tRNA(Asp) confers histidine identity to the transplanted molecule. Optimal expression of histidine identity depends on the chemical nature of the -1 residue and is further increased and/or modulated by the discriminator base N73 and by residues in the anticodon. In the viral tRNA-like domains, the major identity determinant -1 is mimicked by a residue from the single-stranded L1 regions of the different pseudoknots. The consequences of this mimicry for the function of minimalist RNAs derived from tRNA-like domains are discussed. The characteristics of the histidine systems illustrate well the view that the core of the amino acid accepting RNAs is a scaffold that allows proper presentation of identity nucleotides to their amino acid identity counterparts in the synthetase and that different types of scaffoldings are possible.},
note = {0968-0896
Journal Article
Review
Review, Tutorial},
keywords = {Amino Acyl/metabolism RNA, Base Sequence Histidine-tRNA Ligase/*metabolism Molecular Sequence Data RNA, FLORENTZ, Fungal/*metabolism RNA, Non-U.S. Gov't, Transfer, Unité ARN, Viral/metabolism Saccharomyces cerevisiae/*enzymology Substrate Specificity Support},
pubstate = {published},
tppubtype = {article}
}
Histidine aminoacylation systems are of interest because of the structural diversity of the RNA substrates recognized by histidyl-tRNA synthetases. Among tRNAs participating in protein synthesis, those specific for histidine all share an additional residue at their 5'-extremities. On the other hand, tRNA-like domains at the 3'--termini of some plant viruses can also be charged by histidyl-tRNA synthetases, although they are not actors in protein synthesis. This is the case for the RNAs from tobacco mosaic virus and its satellite virus but also those of turnip yellow and brome mosaic viruses. All these RNAs have intricate foldings at their 3'-termini differing from that of canonical tRNAs and share a pseudoknotted domain which is the prerequisite for their folding into structures mimicking the overall L-shape of tRNAs. This paper gives an overview on tRNA identity and rationalizes the apparently contradictory structural and aminoacylation features of histidine-specific tRNAs and tRNA-like structures. The discussion mainly relies on histidylation data obtained with the yeast synthetase, but the conclusions are of a more universal nature. In canonical tRNA(His), the major histidine identity element is the 'minus' 1 residue, since its removal impairs histidylation and conversely its addition to a non-cognate tRNA(Asp) confers histidine identity to the transplanted molecule. Optimal expression of histidine identity depends on the chemical nature of the -1 residue and is further increased and/or modulated by the discriminator base N73 and by residues in the anticodon. In the viral tRNA-like domains, the major identity determinant -1 is mimicked by a residue from the single-stranded L1 regions of the different pseudoknots. The consequences of this mimicry for the function of minimalist RNAs derived from tRNA-like domains are discussed. The characteristics of the histidine systems illustrate well the view that the core of the amino acid accepting RNAs is a scaffold that allows proper presentation of identity nucleotides to their amino acid identity counterparts in the synthetase and that different types of scaffoldings are possible.
Putz J, Wientges J, Sissler M, Giege R, Florentz C, Schwienhorst A
Rapid selection of aminoacyl-tRNAs based on biotinylation of alpha-NH2 group of charged amino acids Article de journal
Dans: Nucleic Acids Res, vol. 25, no. 9, p. 1862-1863, 1997, ISBN: 9162902, (0305-1048 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Amino Acids/*chemistry Biotin/*chemistry Polymerase Chain Reaction RNA, Amino Acyl/chemistry/*isolation & purification Support, FLORENTZ, Non-U.S. Gov't, SISSLER, Transfer, Unité ARN
@article{,
title = {Rapid selection of aminoacyl-tRNAs based on biotinylation of alpha-NH2 group of charged amino acids},
author = {J Putz and J Wientges and M Sissler and R Giege and C Florentz and A Schwienhorst},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9162902},
isbn = {9162902},
year = {1997},
date = {1997-01-01},
journal = {Nucleic Acids Res},
volume = {25},
number = {9},
pages = {1862-1863},
abstract = {A rapid selection procedure to separate low amounts of aminoacylated tRNAs from large pools of inactive variants is described. The procedure involves a three-step protocol. After initial aminoacylation of a tRNA pool, N-hydroxysuccinimide ester chemistry is applied to biotinylate the alpha-NH2 group of the amino acid bound to the 3'-end of a tRNA. The biotin tag is used to capture the derivatized tRNAs on streptavidin-conjugated magnetic beads. Variants bound to the solid phase can be amplified by RT-PCR and transcription, providing tRNAs for subsequent selection rounds.},
note = {0305-1048
Journal Article},
keywords = {Amino Acids/*chemistry Biotin/*chemistry Polymerase Chain Reaction RNA, Amino Acyl/chemistry/*isolation & purification Support, FLORENTZ, Non-U.S. Gov't, SISSLER, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
A rapid selection procedure to separate low amounts of aminoacylated tRNAs from large pools of inactive variants is described. The procedure involves a three-step protocol. After initial aminoacylation of a tRNA pool, N-hydroxysuccinimide ester chemistry is applied to biotinylate the alpha-NH2 group of the amino acid bound to the 3'-end of a tRNA. The biotin tag is used to capture the derivatized tRNAs on streptavidin-conjugated magnetic beads. Variants bound to the solid phase can be amplified by RT-PCR and transcription, providing tRNAs for subsequent selection rounds.
Paillart J C, Westhof E, Ehresmann C, Ehresmann B, Marquet R
Non-canonical interactions in a kissing loop complex: the dimerization initiation site of HIV-1 genomic RNA Article de journal
Dans: J Mol Biol, vol. 270, no. 1, p. 36-49, 1997, ISBN: 9231899, (0022-2836 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Dimerization HIV-1/*genetics Kinetics Models, MARQUET, Molecular Mutagenesis, Non-U.S. Gov't, PAILLART, Site-Directed Nucleic Acid Conformation Purines/chemistry RNA, Unité ARN, Viral/*chemistry/genetics/*metabolism Support
@article{,
title = {Non-canonical interactions in a kissing loop complex: the dimerization initiation site of HIV-1 genomic RNA},
author = {J C Paillart and E Westhof and C Ehresmann and B Ehresmann and R Marquet},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9231899},
isbn = {9231899},
year = {1997},
date = {1997-01-01},
journal = {J Mol Biol},
volume = {270},
number = {1},
pages = {36-49},
abstract = {Retroviruses encapsidate two molecules of genomic RNA that are noncovalently linked close to their 5' ends in a region called the dimer linkage structure (DLS). The dimerization initiation site (DIS) of human immunodeficiency virus type 1 (HIV-1) constitutes the essential part of the DLS in vitro and is crucial for efficient HIV-1 replication in cell culture. We previously identified the DIS as a hairpin structure, located upstream of the major splice donor site, that contains in the loop a six-nucleotide self-complementary sequence preceded and followed by two and one purines, respectively. Two RNA monomers form a kissing loop complex via intermolecular interactions of the six nucleotide self-complementary sequence. Here, we introduced compensatory mutations in the self-complementary sequence and/or a mutation in the flanking purines. We determined the kinetics of dimerization, the thermal stabilities and the apparent equilibrium dissociation constants of wild-type and mutant dimers and used chemical probing to obtain structural information. Our results demonstrate the importance of the 5'-flanking purine and of the two central bases of the self-complementary sequence in the dimerization process. The experimental data are rationalized by triple interactions between these residues in the deep groove of the kissing helix and are incorporated into a three-dimensional model of the kissing loop dimer. In addition, chemical probing and molecular modeling favor the existence of a non-canonical interaction between the conserved adenine residues at the first and last positions in the DIS loop. Furthermore, we show that destabilization of the kissing loop complex at the DIS can be compensated by interactions involving sequences located downstream of the splice donor site of the HIV-1 genomic RNA.},
note = {0022-2836
Journal Article},
keywords = {Dimerization HIV-1/*genetics Kinetics Models, MARQUET, Molecular Mutagenesis, Non-U.S. Gov't, PAILLART, Site-Directed Nucleic Acid Conformation Purines/chemistry RNA, Unité ARN, Viral/*chemistry/genetics/*metabolism Support},
pubstate = {published},
tppubtype = {article}
}
Retroviruses encapsidate two molecules of genomic RNA that are noncovalently linked close to their 5' ends in a region called the dimer linkage structure (DLS). The dimerization initiation site (DIS) of human immunodeficiency virus type 1 (HIV-1) constitutes the essential part of the DLS in vitro and is crucial for efficient HIV-1 replication in cell culture. We previously identified the DIS as a hairpin structure, located upstream of the major splice donor site, that contains in the loop a six-nucleotide self-complementary sequence preceded and followed by two and one purines, respectively. Two RNA monomers form a kissing loop complex via intermolecular interactions of the six nucleotide self-complementary sequence. Here, we introduced compensatory mutations in the self-complementary sequence and/or a mutation in the flanking purines. We determined the kinetics of dimerization, the thermal stabilities and the apparent equilibrium dissociation constants of wild-type and mutant dimers and used chemical probing to obtain structural information. Our results demonstrate the importance of the 5'-flanking purine and of the two central bases of the self-complementary sequence in the dimerization process. The experimental data are rationalized by triple interactions between these residues in the deep groove of the kissing helix and are incorporated into a three-dimensional model of the kissing loop dimer. In addition, chemical probing and molecular modeling favor the existence of a non-canonical interaction between the conserved adenine residues at the first and last positions in the DIS loop. Furthermore, we show that destabilization of the kissing loop complex at the DIS can be compensated by interactions involving sequences located downstream of the splice donor site of the HIV-1 genomic RNA.
Moine H, Nurse K, Ehresmann B, Ehresmann C, Ofengand J
Conformational analysis of Escherichia coli 30S ribosomes containing the single-base mutations G530U, U1498G, G1401C, and C1501G and the double-base mutation G1401C/C1501G Article de journal
Dans: Biochemistry, vol. 36, no. 44, p. 13700-13709, 1997, ISBN: 9354641, (0006-2960 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: 16S/chemical synthesis/*chemistry/genetics Ribosomes/chemistry/genetics Structure-Activity Relationship Support, Bacterial/*chemistry/genetics RNA, Base Sequence Cytosine Nucleotides/genetics Deoxyuridine Escherichia coli/genetics Guanine Nucleotides/genetics Molecular Sequence Data *Mutagenesis, Non-U.S. Gov't, Ribosomal, Site-Directed *Nucleic Acid Conformation RNA, Unité ARN
@article{,
title = {Conformational analysis of Escherichia coli 30S ribosomes containing the single-base mutations G530U, U1498G, G1401C, and C1501G and the double-base mutation G1401C/C1501G},
author = {H Moine and K Nurse and B Ehresmann and C Ehresmann and J Ofengand},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9354641},
isbn = {9354641},
year = {1997},
date = {1997-01-01},
journal = {Biochemistry},
volume = {36},
number = {44},
pages = {13700-13709},
abstract = {Biochemical and genetic studies have pointed out the importance of several sites in 16S ribosomal RNA of Escherichia coli in the decoding process. These sites consist of the core of the decoding center (1400/1500 region) and two other segments (530 and 1050/1200 regions). To detect a possible structural link between these functionally related regions, we analyzed their sensitivity to conformational changes induced by mutations which are located in each of these regions and are known to affect the decoding process. The conformations of five segments of 16S rRNA (1-106, 406-569, 780-978, 997-1247, and 1334-1519) were analyzed by chemical probing of 30S ribosomes containing the following mutations: G530U, U1498G, G1401C, C1501G, and G1401C/C1501G. Ribosomes reconstituted with natural wild-type 16S RNA showed only minor conformational differences with respect to ribosomes isolated from cells. When 16S RNA made in vitro replaced natural 16S RNA, a slightly looser conformation of the central core region was found. Mutant ribosomes made by reconstitution with mutant 16S RNA made in vitro showed conformational effects which were in all cases localized to the region of secondary structure surrounding the site of mutation. Although the core of the decoding center (1400/1500 region) and the two other sites (530 and 1050/1200 regions) participating in the decoding function have been functionally linked, our data indicate that they are structurally independent. They also provide evidence for an unusual structure of the 1400/1500 decoding center, possibly involving noncanonical interactions. Furthermore, the absence of any conformational effect induced by the G530U mutation except at the site of mutation itself points to its direct, as opposed to indirect, involvement in the decoding function of the ribosome.},
note = {0006-2960
Journal Article},
keywords = {16S/chemical synthesis/*chemistry/genetics Ribosomes/chemistry/genetics Structure-Activity Relationship Support, Bacterial/*chemistry/genetics RNA, Base Sequence Cytosine Nucleotides/genetics Deoxyuridine Escherichia coli/genetics Guanine Nucleotides/genetics Molecular Sequence Data *Mutagenesis, Non-U.S. Gov't, Ribosomal, Site-Directed *Nucleic Acid Conformation RNA, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Biochemical and genetic studies have pointed out the importance of several sites in 16S ribosomal RNA of Escherichia coli in the decoding process. These sites consist of the core of the decoding center (1400/1500 region) and two other segments (530 and 1050/1200 regions). To detect a possible structural link between these functionally related regions, we analyzed their sensitivity to conformational changes induced by mutations which are located in each of these regions and are known to affect the decoding process. The conformations of five segments of 16S rRNA (1-106, 406-569, 780-978, 997-1247, and 1334-1519) were analyzed by chemical probing of 30S ribosomes containing the following mutations: G530U, U1498G, G1401C, C1501G, and G1401C/C1501G. Ribosomes reconstituted with natural wild-type 16S RNA showed only minor conformational differences with respect to ribosomes isolated from cells. When 16S RNA made in vitro replaced natural 16S RNA, a slightly looser conformation of the central core region was found. Mutant ribosomes made by reconstitution with mutant 16S RNA made in vitro showed conformational effects which were in all cases localized to the region of secondary structure surrounding the site of mutation. Although the core of the decoding center (1400/1500 region) and the two other sites (530 and 1050/1200 regions) participating in the decoding function have been functionally linked, our data indicate that they are structurally independent. They also provide evidence for an unusual structure of the 1400/1500 decoding center, possibly involving noncanonical interactions. Furthermore, the absence of any conformational effect induced by the G530U mutation except at the site of mutation itself points to its direct, as opposed to indirect, involvement in the decoding function of the ribosome.
Moine H, Cachia C, Westhof E, Ehresmann B, Ehresmann C
The RNA binding site of S8 ribosomal protein of Escherichia coli: Selex and hydroxyl radical probing studies Article de journal
Dans: RNA, vol. 3, no. 3, p. 255-268, 1997, ISBN: 9056763, (1355-8382 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: 16S/chemistry/metabolism Ribosomal Proteins/chemistry/*metabolism Support, Autoradiography Base Sequence Binding Sites Consensus Sequence Escherichia coli Genetic Techniques Hydroxyl Radical/chemistry Models, Bacterial/*metabolism RNA, Molecular Molecular Sequence Data Nucleic Acid Conformation Phylogeny Polymerase Chain Reaction RNA, Non-U.S. Gov't, Ribosomal, Unité ARN
@article{,
title = {The RNA binding site of S8 ribosomal protein of Escherichia coli: Selex and hydroxyl radical probing studies},
author = {H Moine and C Cachia and E Westhof and B Ehresmann and C Ehresmann},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9056763},
isbn = {9056763},
year = {1997},
date = {1997-01-01},
journal = {RNA},
volume = {3},
number = {3},
pages = {255-268},
abstract = {The RNA binding site of ribosomal protein S8 of Escherichia coli is confined to a small region within the stem of a hairpin in 16S rRNA (nt 588-605/633-651), and thus represents a model system for understanding RNA/protein interaction rules. The S8 binding site on 16S rRNA was suspected to contain noncanonical features difficult to prove with classical genetical or biochemical means. We performed in vitro iterative selection of RNA aptamers that bind S8. For the different aptamers, the interactions with the protein were probed with hydroxyl radicals. Aptamers that were recognized according to the same structural rules as wild-type RNA, but with variations not found in nature, were identified. These aptamers revealed features in the S8 binding site that had been concealed during previous characterizations by the high base conservation throughout evolution. Our data demonstrate that the core structure of the S8 binding site is composed of three interdependent bases (nt 597/641/643), with an essential intervening adenine nucleotide (position 642). The other elements important for the binding site are a base pair (598/640) above the three interdependent bases and a bulged base at position 595, the identity of which is not important. Possible implications on the geometry of the S8 binding site are discussed with the help of a three-dimensional model.},
note = {1355-8382
Journal Article},
keywords = {16S/chemistry/metabolism Ribosomal Proteins/chemistry/*metabolism Support, Autoradiography Base Sequence Binding Sites Consensus Sequence Escherichia coli Genetic Techniques Hydroxyl Radical/chemistry Models, Bacterial/*metabolism RNA, Molecular Molecular Sequence Data Nucleic Acid Conformation Phylogeny Polymerase Chain Reaction RNA, Non-U.S. Gov't, Ribosomal, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
The RNA binding site of ribosomal protein S8 of Escherichia coli is confined to a small region within the stem of a hairpin in 16S rRNA (nt 588-605/633-651), and thus represents a model system for understanding RNA/protein interaction rules. The S8 binding site on 16S rRNA was suspected to contain noncanonical features difficult to prove with classical genetical or biochemical means. We performed in vitro iterative selection of RNA aptamers that bind S8. For the different aptamers, the interactions with the protein were probed with hydroxyl radicals. Aptamers that were recognized according to the same structural rules as wild-type RNA, but with variations not found in nature, were identified. These aptamers revealed features in the S8 binding site that had been concealed during previous characterizations by the high base conservation throughout evolution. Our data demonstrate that the core structure of the S8 binding site is composed of three interdependent bases (nt 597/641/643), with an essential intervening adenine nucleotide (position 642). The other elements important for the binding site are a base pair (598/640) above the three interdependent bases and a bulged base at position 595, the identity of which is not important. Possible implications on the geometry of the S8 binding site are discussed with the help of a three-dimensional model.
Masquida B, Felden B, Westhof E
Context dependent RNA-RNA recognition in a three-dimensional model of the 16S rRNA core Article de journal
Dans: Bioorg Med Chem, vol. 5, no. 6, p. 1021-1035, 1997, ISBN: 9222495, (0968-0896 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: 16S/*chemistry/*metabolism Ribosomal Proteins/chemistry Substrate Specificity Support, Base Sequence Escherichia coli/metabolism *Models, Molecular Molecular Sequence Data *Nucleic Acid Conformation Peptide Mapping RNA/*chemistry/*metabolism RNA, Non-U.S. Gov't, Ribosomal, Unité ARN
@article{,
title = {Context dependent RNA-RNA recognition in a three-dimensional model of the 16S rRNA core},
author = {B Masquida and B Felden and E Westhof},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9222495},
isbn = {9222495},
year = {1997},
date = {1997-01-01},
journal = {Bioorg Med Chem},
volume = {5},
number = {6},
pages = {1021-1035},
abstract = {A 3-D model of the core of the 16S rRNA of Escherichia coli containing 328 residues has been built in the protein map derived from neutron scattering data with the help of all the available phylogenetic, biochemical, and cross-linking data. The three pseudoknots of the 16S-core cluster, through the arrangement of complex three-, four- and five-way junctions, around the neck and at the subunit interface. The roles in assembly, initiation or elongation of the three pseudoknots in ribosomal dynamics are emphasized. The 530-loop, localized on the periphery of the 30S particle, could be built with and without a pseudoknot independently of the state of the particle. The pseudoknot of the central domain controls the dynamics of an helix connected to the subunit interface which could trigger some mechanism during translation. The process of the model construction is compatible with a folding scenario in which the 5'-terminal pseudoknot controls the assembly of the central junction and the subsequent folding of the 3'-major domain. The modelling, together with the phylogenetic analysis and the experimental data, point to several potential RNA-RNA contacts which depend on the structural and sequence context in which they occur.},
note = {0968-0896
Journal Article},
keywords = {16S/*chemistry/*metabolism Ribosomal Proteins/chemistry Substrate Specificity Support, Base Sequence Escherichia coli/metabolism *Models, Molecular Molecular Sequence Data *Nucleic Acid Conformation Peptide Mapping RNA/*chemistry/*metabolism RNA, Non-U.S. Gov't, Ribosomal, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
A 3-D model of the core of the 16S rRNA of Escherichia coli containing 328 residues has been built in the protein map derived from neutron scattering data with the help of all the available phylogenetic, biochemical, and cross-linking data. The three pseudoknots of the 16S-core cluster, through the arrangement of complex three-, four- and five-way junctions, around the neck and at the subunit interface. The roles in assembly, initiation or elongation of the three pseudoknots in ribosomal dynamics are emphasized. The 530-loop, localized on the periphery of the 30S particle, could be built with and without a pseudoknot independently of the state of the particle. The pseudoknot of the central domain controls the dynamics of an helix connected to the subunit interface which could trigger some mechanism during translation. The process of the model construction is compatible with a folding scenario in which the 5'-terminal pseudoknot controls the assembly of the central junction and the subsequent folding of the 3'-major domain. The modelling, together with the phylogenetic analysis and the experimental data, point to several potential RNA-RNA contacts which depend on the structural and sequence context in which they occur.
Malmgren C, Wagner E G, Ehresmann C, Ehresmann B, Romby P
Antisense RNA control of plasmid R1 replication. The dominant product of the antisense rna-mrna binding is not a full RNA duplex Article de journal
Dans: J Biol Chem, vol. 272, no. 19, p. 12508-12512, 1997, ISBN: 9139701, (0021-9258 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Antisense/*metabolism RNA, Bacterial Proteins/genetics/metabolism Base Sequence Copper/metabolism Electrophoresis, Messenger/*metabolism Ribonuclease III Support, Non-U.S. Gov't, Polyacrylamide Gel Endoribonucleases/metabolism Escherichia coli Lead Molecular Sequence Data Nucleic Acid Conformation Plasmids/*metabolism Pseudomonas RNA, ROMBY, Unité ARN
@article{,
title = {Antisense RNA control of plasmid R1 replication. The dominant product of the antisense rna-mrna binding is not a full RNA duplex},
author = {C Malmgren and E G Wagner and C Ehresmann and B Ehresmann and P Romby},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9139701},
isbn = {9139701},
year = {1997},
date = {1997-01-01},
journal = {J Biol Chem},
volume = {272},
number = {19},
pages = {12508-12512},
abstract = {The replication frequency of plasmid R1 is controlled by an antisense RNA (CopA) that binds to its target site (CopT) in the leader region of repA mRNA and inhibits the synthesis of the replication initiator protein RepA. Previous studies on CopA-CopT pairing in vitro revealed the existence of a primary loop-loop interaction (kissing complex) that is subsequently converted to an almost irreversible duplex. However, the structure of more stable binding intermediates that lead to the formation of a complete duplex was speculative. Here, we investigated the interaction between CopA and CopT by using Pb(II)-induced cleavages. The kissing complex was studied using a truncated antisense RNA (CopI) that is unable to form a full duplex with CopT. Furthermore, RNase III, which is known to process the CopA-CopT complex in vivo, was used to detect the existence of a full duplex. Our data indicate that the formation of a full CopA-CopT duplex appears to be a very slow process in vitro. Unexpectedly, we found that the loop-loop interaction persists in the predominant CopA-CopT complex and is stabilized by intermolecular base pairing involving the 5'-proximal 30 nucleotides of CopA and the complementary region of CopT. This almost irreversible complex suffices to inhibit ribosome binding at the tap ribosome binding site and may be the inhibitory complex in vivo.},
note = {0021-9258
Journal Article},
keywords = {Antisense/*metabolism RNA, Bacterial Proteins/genetics/metabolism Base Sequence Copper/metabolism Electrophoresis, Messenger/*metabolism Ribonuclease III Support, Non-U.S. Gov't, Polyacrylamide Gel Endoribonucleases/metabolism Escherichia coli Lead Molecular Sequence Data Nucleic Acid Conformation Plasmids/*metabolism Pseudomonas RNA, ROMBY, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
The replication frequency of plasmid R1 is controlled by an antisense RNA (CopA) that binds to its target site (CopT) in the leader region of repA mRNA and inhibits the synthesis of the replication initiator protein RepA. Previous studies on CopA-CopT pairing in vitro revealed the existence of a primary loop-loop interaction (kissing complex) that is subsequently converted to an almost irreversible duplex. However, the structure of more stable binding intermediates that lead to the formation of a complete duplex was speculative. Here, we investigated the interaction between CopA and CopT by using Pb(II)-induced cleavages. The kissing complex was studied using a truncated antisense RNA (CopI) that is unable to form a full duplex with CopT. Furthermore, RNase III, which is known to process the CopA-CopT complex in vivo, was used to detect the existence of a full duplex. Our data indicate that the formation of a full CopA-CopT duplex appears to be a very slow process in vitro. Unexpectedly, we found that the loop-loop interaction persists in the predominant CopA-CopT complex and is stabilized by intermolecular base pairing involving the 5'-proximal 30 nucleotides of CopA and the complementary region of CopT. This almost irreversible complex suffices to inhibit ribosome binding at the tap ribosome binding site and may be the inhibitory complex in vivo.
Isel C, Ehresmann C, Keith G, Ehresmann B, Marquet R
Two step synthesis of (-) strong-stop DNA by avian and murine reverse transcriptases in vitro Article de journal
Dans: Nucleic Acids Res, vol. 25, no. 3, p. 545-552, 1997, ISBN: 9016594, (0305-1048 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Animals Cattle DNA Primers DNA, Avian/*enzymology Rna RNA, MARQUET, Murine/*enzymology Mice Myeloblastosis Virus, Non-U.S. Gov't, Pro RNA, Transfer, Trp RNA-Directed DNA Polymerase/*metabolism Support, Unité ARN, Viral/*biosynthesis Leukemia Virus
@article{,
title = {Two step synthesis of (-) strong-stop DNA by avian and murine reverse transcriptases in vitro},
author = {C Isel and C Ehresmann and G Keith and B Ehresmann and R Marquet},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9016594},
isbn = {9016594},
year = {1997},
date = {1997-01-01},
journal = {Nucleic Acids Res},
volume = {25},
number = {3},
pages = {545-552},
abstract = {Retroviral reverses transcriptases (RTs) are RNA- and DNA-dependent DNA polymerases that use a tRNA bound at the so-called primer binding site (PBS) located near the 5'end of the genomic RNA as primer. Thus, RTs must be able to accommodate both RNA and DNA in the primer strand. To test whether the natural primer confers some advantages to the priming process, we compared initiation of reverse transcription of avian and murine retroviral RNAs, using either their natural tRNA primer, tRNATrp and tRNAPro, respectively, or synthetic 18mer oligodeoxyribonucleotides (ODNs) and oligoribonucleotides (ORNs) complementary to their PBS. In both retroviral systems, the initial extension of ODNs was fast and processive. The initial extension of ORNs, tRNATrp and tRNAPro was much slower and distributive, giving rise to the transient accumulation of short pausing products. Synthesis of (-) strong-stop DNA was delayed when using ORNs and tRNAs, compared to ODNs. Even though ORNs and tRNAs were initially extended at the same rate, the short pausing products were more rapidly extended when using the tRNA primers. As a consequence, synthesis of (-) strong-stop DNA was much more efficient with tRNA primers, compared to ORNs. Taken together, these results suggest that the tRNA-primed synthesis of (-) strong-stop DNA is a two-step process, as already observed for HIV-1. The initiation mode corresponds to the initial non-processive nucleotide addition and extension of the short pausing products. It is more efficient with the natural primers than with ORNs. Initiation is followed by a more processive and unspecific elongation mode. Elongation is observed when the primer strand is DNA, i.e. when using the ODNs as primers or when the ORN and tRNA primers have been extended by a sufficient number (depending on the retroviral system) of deoxyribonucleotides.},
note = {0305-1048
Journal Article},
keywords = {Animals Cattle DNA Primers DNA, Avian/*enzymology Rna RNA, MARQUET, Murine/*enzymology Mice Myeloblastosis Virus, Non-U.S. Gov't, Pro RNA, Transfer, Trp RNA-Directed DNA Polymerase/*metabolism Support, Unité ARN, Viral/*biosynthesis Leukemia Virus},
pubstate = {published},
tppubtype = {article}
}