Publications
2003
Rigourd M, Goldschmidt V, Brule F, Morrow C D, Ehresmann B, Ehresmann C, Marquet R
Structure-function relationships of the initiation complex of HIV-1 reverse transcription: the case of mutant viruses using tRNA(His) as primer Journal Article
In: Nucleic Acids Res, vol. 31, no. 19, pp. 5764-5775, 2003, ISBN: 14500840, (1362-4962 Journal Article).
Abstract | Links | BibTeX | Tags: Base Sequence Comparative Study DNA, Genetic, Genetic *Transcription Initiation Site *Transcription, His/*metabolism RNA, MARQUET, Non-U.S. Gov't Templates, Post-Transcriptional RNA, Transfer, Unité ARN, Viral HIV-1/*genetics/metabolism HIV-1 Reverse Transcriptase/metabolism Kinetics Macromolecular Systems Molecular Sequence Data Mutation RNA Probes RNA Processing, Viral/*biosynthesis/genetics Sequence Alignment Structure-Activity Relationship Support, Viral/biosynthesis *Gene Expression Regulation
@article{,
title = {Structure-function relationships of the initiation complex of HIV-1 reverse transcription: the case of mutant viruses using tRNA(His) as primer},
author = {M Rigourd and V Goldschmidt and F Brule and C D Morrow and B Ehresmann and C Ehresmann and R Marquet},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14500840},
isbn = {14500840},
year = {2003},
date = {2003-01-01},
journal = {Nucleic Acids Res},
volume = {31},
number = {19},
pages = {5764-5775},
abstract = {Reverse transcription of HIV-1 RNA is initiated from the 3' end of a tRNA3Lys molecule annealed to the primer binding site (PBS). An additional interaction between the anticodon loop of tRNA3Lys and a viral A-rich loop is required for efficient initiation of reverse transcription of the HIV-1 MAL isolate. In the HIV-1 HXB2 isolate, simultaneous mutations of the PBS and the A-rich loop (mutant His-AC), but not of the PBS alone (mutant His) allows the virus to stably utilize tRNA(His) as primer. However, mutant His-AC selects additional mutations during cell culture, generating successively His-AC-GAC and His-AC-AT-GAC. Here, we wanted to establish direct relationships between the evolution of these mutants in cell culture, their efficiency in initiating reverse transcription and the structure of the primer/template complexes in vitro. The initiation of reverse transcription of His and His-AC RNAs was dramatically reduced. However, His-AC-GAC RNA, which incorporated three adaptative point mutations, was reverse transcribed more efficiently than the wild type RNA. Incorporation of two additional mutations decreased the efficiency of the initiation of reverse transcription, which remained at the wild type level. Structural probing showed that even though both His-AC and His-AC-GAC RNAs can potentially interact with the anticodon loop of tRNA(His), only the latter template formed a stable interaction. Thus, our results showed that the selection of adaptative mutations by HIV-1 mutants utilizing tRNA(His) as primer was initially dictated by the efficiency of the initiation of reverse transcription, which relied on the existence of a stable interaction between the mutated A-rich loop and the anticodon loop of tRNA(His).},
note = {1362-4962
Journal Article},
keywords = {Base Sequence Comparative Study DNA, Genetic, Genetic *Transcription Initiation Site *Transcription, His/*metabolism RNA, MARQUET, Non-U.S. Gov't Templates, Post-Transcriptional RNA, Transfer, Unité ARN, Viral HIV-1/*genetics/metabolism HIV-1 Reverse Transcriptase/metabolism Kinetics Macromolecular Systems Molecular Sequence Data Mutation RNA Probes RNA Processing, Viral/*biosynthesis/genetics Sequence Alignment Structure-Activity Relationship Support, Viral/biosynthesis *Gene Expression Regulation},
pubstate = {published},
tppubtype = {article}
}
2002
Brule F, Marquet R, Rong L, Wainberg M A, Roques B P, Grice S F Le, Ehresmann B, Ehresmann C
In: RNA, vol. 8, no. 1, pp. 8-15, 2002, ISBN: 11873759, (1355-8382 Journal Article).
Abstract | Links | BibTeX | Tags: Genetic, Genetic Transcription, Lys/*chemistry/genetics/metabolism RNA, MARQUET, Non-U.S. Gov't Templates, Transfer, Unité ARN, Viral/*chemistry/genetics/metabolism Support
@article{,
title = {Structural and functional properties of the HIV-1 RNA-tRNA(Lys)3 primer complex annealed by the nucleocapsid protein: comparison with the heat-annealed complex},
author = {F Brule and R Marquet and L Rong and M A Wainberg and B P Roques and S F Le Grice and B Ehresmann and C Ehresmann},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11873759},
isbn = {11873759},
year = {2002},
date = {2002-01-01},
journal = {RNA},
volume = {8},
number = {1},
pages = {8-15},
abstract = {The conversion of the single-stranded RNA genome into double-stranded DNA by virus-coded reverse transcriptase (RT) is an essential step of the retrovirus life cycle. In human immunodeficiency virus type 1 (HIV-1), RT uses the cellular tRNA(Lys)3 to initiate the (-) strand DNA synthesis. Placement of the primer tRNA(Lys)3 involves binding of its 3'-terminal 18 nt to a complementary region of genomic RNA termed PBS. However, the PBS sequence is not the unique determinant of primer usage and additional contacts are important. This placement is believed to be achieved in vivo by the nucleocapsid domain of Gag or by the mature protein NCp. Up to now, structural information essentially arose from heat-annealed primer-template complexes (Isel et al., J Mol Biol, 1995, 247:236-250; Isel et al., EMBO J, 1999, 18:1038-1048). Here, we investigated the formation of the primer-template complex mediated by NCp and compared structural and functional properties of heat- and NCp-annealed complexes. We showed that both heat- and NCp-mediated procedures allow comparable high yields of annealing. Then, we investigated structural features of both kinds of complexes by enzymatic probing, and we compared their relative efficiency in (-) strong stop DNA synthesis. We did not find any significant differences between these complexes, suggesting that information derived from the heat-annealed complex can be transposed to the NCp-mediated complex and most likely to complexes formed in vivo.},
note = {1355-8382
Journal Article},
keywords = {Genetic, Genetic Transcription, Lys/*chemistry/genetics/metabolism RNA, MARQUET, Non-U.S. Gov't Templates, Transfer, Unité ARN, Viral/*chemistry/genetics/metabolism Support},
pubstate = {published},
tppubtype = {article}
}
2000
Wilhelm M, Boutabout M, Wilhelm F X
In: Biochem J, vol. 348, no. Pt 2, pp. 337-342, 2000, ISBN: 10816427, (0264-6021 Journal Article).
Abstract | Links | BibTeX | Tags: Affinity Cloning, Amino Acid Support, Amino Acid Sequence Chromatography, Calf Thymus/isolation & purification/*metabolism Saccharomyces cerevisiae/*enzymology/*genetics Sequence Alignment Sequence Homology, Genetic, Molecular Codon, Non-U.S. Gov't Templates, Unité ARN
@article{,
title = {Expression of an active form of recombinant Ty1 reverse transcriptase in Escherichia coli: a fusion protein containing the C-terminal region of the Ty1 integrase linked to the reverse transcriptase-RNase H domain exhibits polymerase and RNase H activities},
author = {M Wilhelm 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=10816427},
isbn = {10816427},
year = {2000},
date = {2000-01-01},
journal = {Biochem J},
volume = {348},
number = {Pt 2},
pages = {337-342},
abstract = {Replication of the Saccharomyces cerevisiae Ty1 retrotransposon requires a reverse transcriptase capable of synthesizing Ty1 DNA. The first description of an active form of a recombinant Ty1 enzyme with polymerase and RNase H activities is reported here. The Ty1 enzyme was expressed as a hexahistidine-tagged fusion protein in Escherichia coli to facilitate purification of the recombinant protein by metal-chelate chromatography. Catalytic activity of the recombinant protein was detected only when amino acid residues encoded by the integrase gene were added to the N-terminus of the reverse transcriptase-RNase H domain. This suggests that the integrase domain could play a role in proper folding of reverse transcriptase. Several biochemical properties of the Ty1 enzyme were analysed, including the effect of MgCl(2), NaCl, temperature and of the chain terminator dideoxy GTP on its polymerase activity. RNase H activity was examined by monitoring the cleavage of a RNA-DNA template-primer. Our results suggest that the distance between the RNase H and polymerase active sites corresponds to the length of a 14-nucleotide RNA-DNA heteroduplex. The recombinant protein produced in E. coli should be useful for further biochemical and structural analyses and for a better understanding of the role of integrase in the activation of reverse transcriptase.},
note = {0264-6021
Journal Article},
keywords = {Affinity Cloning, Amino Acid Support, Amino Acid Sequence Chromatography, Calf Thymus/isolation & purification/*metabolism Saccharomyces cerevisiae/*enzymology/*genetics Sequence Alignment Sequence Homology, Genetic, Molecular Codon, Non-U.S. Gov't Templates, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Tisne C, Rigourd M, Marquet R, Ehresmann C, Dardel F
In: RNA, vol. 6, no. 10, pp. 1403-1412, 2000, ISBN: 11073216, (1355-8382 Journal Article).
Abstract | Links | BibTeX | Tags: Base Sequence DNA, Biomolecular *Nucleic Acid Conformation RNA/*chemistry/genetics/metabolism RNA, Genetic Transcription, Genetic/*genetics Virus Replication, Lys/*chemistry/genetics/metabolism Structure-Activity Relationship Support, MARQUET, Molecular Molecular Sequence Data Mutation/genetics Nuclear Magnetic Resonance, Non-U.S. Gov't Templates, Transfer, Unité ARN, Viral/biosynthesis/genetics Escherichia coli/genetics *Genetic Engineering HIV-1/*genetics/physiology Human Iodine/metabolism Models
@article{,
title = {NMR and biochemical characterization of recombinant human tRNA(Lys)3 expressed in Escherichia coli: identification of posttranscriptional nucleotide modifications required for efficient initiation of HIV-1 reverse transcription},
author = {C Tisne and M Rigourd and R Marquet and C Ehresmann and F Dardel},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11073216},
isbn = {11073216},
year = {2000},
date = {2000-01-01},
journal = {RNA},
volume = {6},
number = {10},
pages = {1403-1412},
abstract = {Reverse transcription of HIV-1 viral RNA uses human tRNA(Lys)3 as a primer. Some of the modified nucleotides carried by this tRNA must play a key role in the initiation of this process, because unmodified tRNA produced in vitro is only marginally active as primer. To provide a better understanding of the contribution of base modifications in the initiation complex, we have designed a recombinant system that allows tRNA(Lys)3 expression in Escherichia coli. Because of their high level of overexpression, some modifications are incorporated at substoichiometric levels. We have purified the two major recombinant tRNA(Lys)3 subspecies, and their modified nucleotide contents have been characterized by a combination of NMR and biochemical techniques. Both species carry psis, Ds, T, t6A, and m7G. Differences are observed at position 34, within the anticodon. One fraction lacks the 5-methylaminomethyl group, whereas the other lacks the 2-thio group. Although the s2U34-containing recombinant tRNA is a less efficient primer, it presents most of the characteristics of the mammalian tRNA. On the other hand, the mnm5U34-containing tRNA has a strongly reduced activity. Our results demonstrate that the modifications that are absent in E. coli (m2G10, psi27, m5C48, m5C49, and m1A58) as well as the mnm5 group at position 34 are dispensable for initiation of reverse transcription. In contrast, the 2-thio group at position 34 seems to play an important part in this process.},
note = {1355-8382
Journal Article},
keywords = {Base Sequence DNA, Biomolecular *Nucleic Acid Conformation RNA/*chemistry/genetics/metabolism RNA, Genetic Transcription, Genetic/*genetics Virus Replication, Lys/*chemistry/genetics/metabolism Structure-Activity Relationship Support, MARQUET, Molecular Molecular Sequence Data Mutation/genetics Nuclear Magnetic Resonance, Non-U.S. Gov't Templates, Transfer, Unité ARN, Viral/biosynthesis/genetics Escherichia coli/genetics *Genetic Engineering HIV-1/*genetics/physiology Human Iodine/metabolism Models},
pubstate = {published},
tppubtype = {article}
}
Schaub M, Krol A, Carbon P
Structural organization of Staf-DNA complexes Journal Article
In: Nucleic Acids Res, vol. 28, no. 10, pp. 2114-2121, 2000, ISBN: 10773080, (1362-4962 Journal Article).
Abstract | Links | BibTeX | Tags: Amino Acid Sequence Animals Base Sequence Binding Sites DNA-Binding Proteins/*chemistry/*metabolism Human Models, Amino Acid-Specific/*genetics Support, Genetic Trans-Activators/*chemistry/*metabolism Vertebrates Xenopus laevis Zinc Fingers, Molecular Molecular Sequence Data Nucleic Acid Conformation Plasmids/*chemistry/*metabolism Protein Conformation RNA, Non-U.S. Gov't Templates, Small Nuclear/*genetics RNA, Transfer, Unité ARN
@article{,
title = {Structural organization of Staf-DNA complexes},
author = {M Schaub and A Krol and P Carbon},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10773080},
isbn = {10773080},
year = {2000},
date = {2000-01-01},
journal = {Nucleic Acids Res},
volume = {28},
number = {10},
pages = {2114-2121},
abstract = {The transactivator Staf, which contains seven contiguous zinc fingers of the C(2)-H(2)type, exerts its effects on gene expression by binding to specific targets in vertebrate small nuclear RNA (snRNA) and snRNA-type gene promoters. Here, we have investigated the interaction of the Staf zinc finger domain with the optimal Xenopus selenocysteine tRNA (xtRNA(Sec)) and human U6 snRNA (hU6) Staf motifs. Generation of a series of polypeptides containing increasing numbers of Staf zinc fingers tested in binding assays, by interference techniques and by binding site selection served to elucidate the mode of interaction between the zinc fingers and the Staf motifs. Our results provide strong evidence that zinc fingers 3-6 represent the minimal zinc finger region for high affinity binding to Staf motifs. Furthermore, we show that the binding of Staf is achieved through a broad spectrum of close contacts between zinc fingers 1-6 and xtRNA(Sec)or optimal sites or between zinc fingers 3-6 and the hU6 site. Extensive DNA major groove contacts contribute to the interaction with Staf that associates more closely with the non-template than with the template strand. Based on these findings and the structural information provided by the solved structures of other zinc finger-DNA complexes, we propose a model for the interaction between Staf zinc fingers and the xtRNA(Sec), optimal and hU6 sites.},
note = {1362-4962
Journal Article},
keywords = {Amino Acid Sequence Animals Base Sequence Binding Sites DNA-Binding Proteins/*chemistry/*metabolism Human Models, Amino Acid-Specific/*genetics Support, Genetic Trans-Activators/*chemistry/*metabolism Vertebrates Xenopus laevis Zinc Fingers, Molecular Molecular Sequence Data Nucleic Acid Conformation Plasmids/*chemistry/*metabolism Protein Conformation RNA, Non-U.S. Gov't Templates, Small Nuclear/*genetics RNA, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Lanchy J M, Isel C, Keith G, Grice S F Le, Ehresmann C, Ehresmann B, Marquet R
Dynamics of the HIV-1 reverse transcription complex during initiation of DNA synthesis Journal Article
In: J Biol Chem, vol. 275, no. 16, pp. 12306-12312, 2000, ISBN: 10766870, (0021-9258 Journal Article).
Abstract | Links | BibTeX | Tags: *Anticodon Base Sequence *DNA Replication *Hiv-1 Human Molecular Sequence Data Mutagenesis Nucleic Acid Conformation RNA, Genetic, Lys/genetics/metabolism RNA, MARQUET, Non-U.S. Gov't Templates, Transfer, Unité ARN, Viral/genetics/metabolism RNA-Directed DNA Polymerase/*metabolism Support
@article{,
title = {Dynamics of the HIV-1 reverse transcription complex during initiation of DNA synthesis},
author = {J M Lanchy and C Isel and G Keith and S F Le Grice 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=10766870},
isbn = {10766870},
year = {2000},
date = {2000-01-01},
journal = {J Biol Chem},
volume = {275},
number = {16},
pages = {12306-12312},
abstract = {Initiation of human immunodeficiency virus-1 (HIV-1) reverse transcription requires formation of a complex containing the viral RNA (vRNA), tRNA(3)(Lys) and reverse transcriptase (RT). The vRNA and the primer tRNA(3)(Lys) form several intermolecular interactions in addition to annealing of the primer 3' end to the primer binding site (PBS). These interactions are crucial for the efficiency and the specificity of the initiation of reverse transcription. However, as they are located upstream of the PBS, they must unwind as DNA synthesis proceeds. Here, the dynamics of the complex during initiation of reverse transcription was followed by enzymatic probing. Our data revealed reciprocal effects of the tertiary structure of the vRNA.tRNA(3)(Lys) complex and reverse transcriptase (RT) at a distance from the polymerization site. The structure of the initiation complex allowed RT to interact with the template strand up to 20 nucleotides upstream from the polymerization site. Conversely, nucleotide addition by RT modified the tertiary structure of the complex at 10-14 nucleotides from the catalytic site. The viral sequences became exposed at the surface of the complex as they dissociated from the tRNA following primer extension. However, the counterpart tRNA sequences became buried inside the complex. Surprisingly, they became exposed when mutations prevented the intermolecular interactions in the initial complex, indicating that the fate of the tRNA depended on the tertiary structure of the initial complex.},
note = {0021-9258
Journal Article},
keywords = {*Anticodon Base Sequence *DNA Replication *Hiv-1 Human Molecular Sequence Data Mutagenesis Nucleic Acid Conformation RNA, Genetic, Lys/genetics/metabolism RNA, MARQUET, Non-U.S. Gov't Templates, Transfer, Unité ARN, Viral/genetics/metabolism RNA-Directed DNA Polymerase/*metabolism Support},
pubstate = {published},
tppubtype = {article}
}
1999
Schaub M, Myslinski E, Krol A, Carbon P
Maximization of selenocysteine tRNA and U6 small nuclear RNA transcriptional activation achieved by flexible utilization of a Staf zinc finger Journal Article
In: J Biol Chem, vol. 274, no. 35, pp. 25042-25050, 1999, ISBN: 10455183, (0021-9258 Journal Article).
Abstract | Links | BibTeX | Tags: Amino Acid Sequence Animals Binding Sites/genetics DNA/genetics DNA-Binding Proteins/*genetics Human Hydroxyl Radical/metabolism Microinjections Molecular Sequence Data Mutation Oocytes Peptide Fragments/immunology Promoter Regions (Genetics) Protein Binding RNA, Amino Acid-Specific/*genetics Sequence Homology Support, Genetic Trans-Activation (Genetics)/*genetics Trans-Activators/*genetics Transcription Factors/genetics Xenopus Zinc Fingers/*genetics, Non-U.S. Gov't Templates, Small Nuclear/*genetics RNA, Transfer, Unité ARN
@article{,
title = {Maximization of selenocysteine tRNA and U6 small nuclear RNA transcriptional activation achieved by flexible utilization of a Staf zinc finger},
author = {M Schaub and E Myslinski and A Krol and P Carbon},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10455183},
isbn = {10455183},
year = {1999},
date = {1999-01-01},
journal = {J Biol Chem},
volume = {274},
number = {35},
pages = {25042-25050},
abstract = {Transcriptional activators Staf and Oct-1 play critical roles in the activation of small nuclear RNA (snRNA) and snRNA-type gene transcription. Recently, we established that Staf binding to the human U6 snRNA (hU6) and Xenopus selenocysteine tRNA (xtRNA(Sec)) genes requires different sets of the seven C2-H2 zinc fingers. In this work, using a combination of oocyte microinjection, electrophoretic mobility shift assays, and missing nucleoside experiments with wild-type and mutant promoters, we demonstrate that the hU6 gene requires zinc fingers 2-7 for Staf binding and Oct-1 for maximal transcriptional activity. In contrast, the xtRNA(Sec) gene needs the binding of the seven Staf zinc fingers, but not Oct-1, for optimal transcriptional capacity. Mutation in the binding site for Staf zinc finger 1 in the tRNA(Sec) promoter reduced both Staf binding and transcriptional activity. Conversely, introduction of a zinc finger 1 binding site in the hU6 promoter increased Staf binding but interfered with the simultaneous Staf and Oct-1 binding, thus reducing transcriptional activity. Collectively, these results show that the differential utilization of Staf zinc finger 1 represents a new, critical determinant of the transcriptional activation mechanism for the Xenopus tRNA(Sec) and human U6 snRNA genes.},
note = {0021-9258
Journal Article},
keywords = {Amino Acid Sequence Animals Binding Sites/genetics DNA/genetics DNA-Binding Proteins/*genetics Human Hydroxyl Radical/metabolism Microinjections Molecular Sequence Data Mutation Oocytes Peptide Fragments/immunology Promoter Regions (Genetics) Protein Binding RNA, Amino Acid-Specific/*genetics Sequence Homology Support, Genetic Trans-Activation (Genetics)/*genetics Trans-Activators/*genetics Transcription Factors/genetics Xenopus Zinc Fingers/*genetics, Non-U.S. Gov't Templates, Small Nuclear/*genetics RNA, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
1996
Isel C, Lanchy J M, Grice S F Le, Ehresmann C, Ehresmann B, Marquet R
In: EMBO J, vol. 15, no. 4, pp. 917-924, 1996, ISBN: 8631312, (0261-4189 Journal Article).
Abstract | Links | BibTeX | Tags: Cell-Free System Gene Expression Regulation, Genetic *Virus Replication, Lys/*metabolism RNA, MARQUET, Non-U.S. Gov't Templates, Transfer, Unité ARN, Viral HIV-1/*genetics HIV-1 Reverse Transcriptase RNA, Viral/metabolism RNA-Directed DNA Polymerase/*metabolism Support
@article{,
title = {Specific initiation and switch to elongation of human immunodeficiency virus type 1 reverse transcription require the post-transcriptional modifications of primer tRNA3Lys},
author = {C Isel and J M Lanchy and S F Le Grice 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=8631312},
isbn = {8631312},
year = {1996},
date = {1996-01-01},
journal = {EMBO J},
volume = {15},
number = {4},
pages = {917-924},
abstract = {Initiation of RNA-dependent DNA synthesis by retroviral reverse transcriptases is generally considered as unspecific. In the case of human immunodeficiency virus type 1 (HIV-1), the natural primer is tRNA3Lys. We recently found evidence of complex interactions between tRNA3Lys and HIV-1 RNA that may be involved in the priming process. In this study, we compare the ability of natural and unmodified synthetic tRNA3Lys and 18mer oligoribo- and oligodeoxyribonucleotides complementary to the viral primer binding site to initiate replication of HIV-1 RNA using either homologous or heterologous reverse transcriptases. We show that HIV-1 RNA, HIV-1 reverse transcriptase and primer tRNA3Lys form a specific initiation complex that differs from the unspecific elongation complex formed when an oligodeoxyribonucleotide is used as primer. Modified nucleosides of tRNA3Lys are required for efficient initiation and transition to elongation. Transition from initiation to elongation, but not initiation of reverse transcription itself, is facilitated by extended primer-template interactions. Elongation, but not initiation of reverse transcription, is inhibited by Mn2+, which further differentiates these two different functional states of reverse transcriptase. These results define initiation of reverse transcription as a target to block viral replication.},
note = {0261-4189
Journal Article},
keywords = {Cell-Free System Gene Expression Regulation, Genetic *Virus Replication, Lys/*metabolism RNA, MARQUET, Non-U.S. Gov't Templates, Transfer, Unité ARN, Viral HIV-1/*genetics HIV-1 Reverse Transcriptase RNA, Viral/metabolism RNA-Directed DNA Polymerase/*metabolism Support},
pubstate = {published},
tppubtype = {article}
}
1994
Frugier M, Florentz C, Hosseini M W, Lehn J M, Giege R
Synthetic polyamines stimulate in vitro transcription by T7 RNA polymerase Journal Article
In: Nucleic Acids Res, vol. 22, no. 14, pp. 2784-2790, 1994, ISBN: 8052534, (0305-1048 Journal Article).
Abstract | Links | BibTeX | Tags: Bacteriophage T7/enzymology Base Sequence Comparative Study DNA-Directed RNA Polymerases/drug effects/*metabolism Kinetics Molecular Sequence Data Molecular Structure Nucleic Acid Conformation Oligodeoxyribonucleotides Polyamines/chemistry/*pharmacology Promoter Regions (Genetics) RNA, ERIANI, FLORENTZ, FRUGIER, Genetic Transcription, Genetic/*drug effects, Non-U.S. Gov't Templates, Transfer, Unité ARN, Val/*biosynthesis/chemistry Structure-Activity Relationship Support
@article{,
title = {Synthetic polyamines stimulate in vitro transcription by T7 RNA polymerase},
author = {M Frugier and C Florentz and M W Hosseini and J M Lehn and R Giege},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8052534},
isbn = {8052534},
year = {1994},
date = {1994-01-01},
journal = {Nucleic Acids Res},
volume = {22},
number = {14},
pages = {2784-2790},
abstract = {The influence of nine synthetic polyamines on in vitro transcription with T7 RNA polymerase has been studied. The compounds used were linear or macrocyclic tetra- and hexaamine, varying in their size, shape and number of protonated groups. Their effect was tested on different types of templates, all presenting the T7 RNA promoter in a double-stranded form followed by sequences encoding short transcripts (25 to 35-mers) either on single- or double-stranded synthetic oligodeoxyribonucleotides. All polyamines used stimulate transcription of both types of templates at levels dependent on their size, shape, protonation degree, and concentration. For each compound, an optimal concentration could be defined; above this concentration, transcription inhibition occurred. Highest stimulation (up to 12-fold) was obtained by the largest cyclic compound called [38]N6C10.},
note = {0305-1048
Journal Article},
keywords = {Bacteriophage T7/enzymology Base Sequence Comparative Study DNA-Directed RNA Polymerases/drug effects/*metabolism Kinetics Molecular Sequence Data Molecular Structure Nucleic Acid Conformation Oligodeoxyribonucleotides Polyamines/chemistry/*pharmacology Promoter Regions (Genetics) RNA, ERIANI, FLORENTZ, FRUGIER, Genetic Transcription, Genetic/*drug effects, Non-U.S. Gov't Templates, Transfer, Unité ARN, Val/*biosynthesis/chemistry Structure-Activity Relationship Support},
pubstate = {published},
tppubtype = {article}
}
1993
Isel C, Marquet R, Keith G, Ehresmann C, Ehresmann B
Modified nucleotides of tRNA(3Lys) modulate primer/template loop-loop interaction in the initiation complex of HIV-1 reverse transcription Journal Article
In: J Biol Chem, vol. 268, no. 34, pp. 25269-25272, 1993, ISBN: 7503978, (0021-9258 Journal Article).
Abstract | Links | BibTeX | Tags: Anticodon/genetics/metabolism Base Sequence Binding Sites Comparative Study DNA Primers HIV-1/genetics/*metabolism HIV-1 Reverse Transcriptase HIV-2/genetics/metabolism Molecular Sequence Data Nucleic Acid Conformation RNA, Genetic, Genetic *Transcription, Lys/*metabolism RNA, MARQUET, Non-U.S. Gov't Templates, Transfer, Unité ARN, Viral/*biosynthesis RNA-Directed DNA Polymerase/*metabolism SIV/genetics/metabolism Support
@article{,
title = {Modified nucleotides of tRNA(3Lys) modulate primer/template loop-loop interaction in the initiation complex of HIV-1 reverse transcription},
author = {C Isel and R Marquet and G Keith and C Ehresmann and B Ehresmann},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=7503978},
isbn = {7503978},
year = {1993},
date = {1993-01-01},
journal = {J Biol Chem},
volume = {268},
number = {34},
pages = {25269-25272},
abstract = {In all retroviruses, reverse transcription is primed by a tRNA whose 3' end 18 nucleotides are complementary to the so called viral primer binding site. Previous work showed that reverse transcription of HIV-1 RNA is initiated by tRNA(3Lys). Using a variety of chemical and enzymatic structural probes, we investigated the interactions between HIV-1 RNA and its natural primer tRNA(3Lys). In addition to the predictable contacts between the viral primer binding site and the 3' end of tRNA(3Lys), a specific interaction takes place between an A-rich loop located upstream of the primer binding site region and the anticodon loop of tRNA(3Lys). This AAAA/Umcm5s2UUU loop-loop interaction is not observed when the natural primer is replaced by an in vitro synthesized tRNA(3Lys) transcript. Furthermore, dethiolation of the modified nucleotide mcm5s2U at position 34 of tRNA(3Lys) strongly destabilizes this interaction. Sequence and structure comparisons indicate that the primer/template loop-loop interaction is conserved in all HIV-1 isolates, and possibly also in HIV-2 and SIV.},
note = {0021-9258
Journal Article},
keywords = {Anticodon/genetics/metabolism Base Sequence Binding Sites Comparative Study DNA Primers HIV-1/genetics/*metabolism HIV-1 Reverse Transcriptase HIV-2/genetics/metabolism Molecular Sequence Data Nucleic Acid Conformation RNA, Genetic, Genetic *Transcription, Lys/*metabolism RNA, MARQUET, Non-U.S. Gov't Templates, Transfer, Unité ARN, Viral/*biosynthesis RNA-Directed DNA Polymerase/*metabolism SIV/genetics/metabolism Support},
pubstate = {published},
tppubtype = {article}
}