Simonetti A, Marzi S, Billas I M, Tsai A, Fabbretti A, Myasnikov A G, Roblin P, Vaiana A C, Hazemann I, Eiler D, Steitz T A, Puglisi J D, Gualerzi C O, Klaholz B P
Involvement of protein IF2 N domain in ribosomal subunit joining revealed from architecture and function of the full-length initiation factor. Article de journal
Dans: Proc Natl Acad Sci U S A, vol. 110, no. 39, p. 15656-61, 2013, ISBN: 24029017.
Résumé | Liens | BibTeX | Étiquettes: ERIANI, integrated structural biology protein synthesis, ROMBY, Unité ARN
@article{,
title = {Involvement of protein IF2 N domain in ribosomal subunit joining revealed from architecture and function of the full-length initiation factor.},
author = {A Simonetti and S Marzi and I M Billas and A Tsai and A Fabbretti and A G Myasnikov and P Roblin and A C Vaiana and I Hazemann and D Eiler and T A Steitz and J D Puglisi and C O Gualerzi and B P Klaholz},
url = {http://www.ncbi.nlm.nih.gov/pubmed/24029017?dopt=Abstract},
doi = {10.1073/pnas.1309578110},
isbn = {24029017},
year = {2013},
date = {2013-01-01},
journal = {Proc Natl Acad Sci U S A},
volume = {110},
number = {39},
pages = {15656-61},
abstract = {Translation initiation factor 2 (IF2) promotes 30S initiation complex (IC) formation and 50S subunit joining, which produces the 70S IC. The architecture of full-length IF2, determined by small angle X-ray diffraction and cryo electron microscopy, reveals a more extended conformation of IF2 in solution and on the ribosome than in the crystal. The N-terminal domain is only partially visible in the 30S IC, but in the 70S IC, it stabilizes interactions between IF2 and the L7/L12 stalk of the 50S, and on its deletion, proper N-formyl-methionyl(fMet)-tRNAfMet positioning and efficient transpeptidation are affected. Accordingly, fast kinetics and single-molecule fluorescence data indicate that the N terminus promotes 70S IC formation by stabilizing the productive sampling of the 50S subunit during 30S IC joining. Together, our data highlight the dynamics of IF2-dependent ribosomal subunit joining and the role played by the N terminus of IF2 in this process.},
keywords = {ERIANI, integrated structural biology protein synthesis, ROMBY, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Simonetti A, Marzi S, Fabbretti A, Hazemann I, Jenner L, Urzhumtsev A, Gualerzi C O, Klaholz B P
Structure of the protein core of translation initiation factor 2 in apo, GTP-bound and GDP-bound forms. Article de journal
Dans: Acta Crystallogr D Biol Crystallogr, vol. 69, no. Pt 6, p. 925-33, 2013, ISBN: 23695237.
Résumé | Liens | BibTeX | Étiquettes: ERIANI, ROMBY, Unité ARN
@article{,
title = {Structure of the protein core of translation initiation factor 2 in apo, GTP-bound and GDP-bound forms.},
author = {A Simonetti and S Marzi and A Fabbretti and I Hazemann and L Jenner and A Urzhumtsev and C O Gualerzi and B P Klaholz},
url = {http://www.ncbi.nlm.nih.gov/pubmed/23695237?dopt=Abstract},
doi = {10.1107/S0907444913006422},
isbn = {23695237},
year = {2013},
date = {2013-01-01},
journal = {Acta Crystallogr D Biol Crystallogr},
volume = {69},
number = {Pt 6},
pages = {925-33},
abstract = {Translation initiation factor 2 (IF2) is involved in the early steps of bacterial protein synthesis. It promotes the stabilization of the initiator tRNA on the 30S initiation complex (IC) and triggers GTP hydrolysis upon ribosomal subunit joining. While the structure of an archaeal homologue (a/eIF5B) is known, there are significant sequence and functional differences in eubacterial IF2, while the trimeric eukaryotic IF2 is completely unrelated. Here, the crystal structure of the apo IF2 protein core from Thermus thermophilus has been determined by MAD phasing and the structures of GTP and GDP complexes were also obtained. The IF2-GTP complex was trapped by soaking with GTP in the cryoprotectant. The structures revealed conformational changes of the protein upon nucleotide binding, in particular in the P-loop region, which extend to the functionally relevant switch II region. The latter carries a catalytically important and conserved histidine residue which is observed in different conformations in the GTP and GDP complexes. Overall, this work provides the first crystal structure of a eubacterial IF2 and suggests that activation of GTP hydrolysis may occur by a conformational repositioning of the histidine residue.},
keywords = {ERIANI, ROMBY, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Martin F
Fifteen years of the yeast three-hybrid system: RNA-protein interactions under investigation. Article de journal
Dans: Methods, vol. 58, no. 4, p. 367-375, 2012, ISBN: 2284156.
Résumé | Liens | BibTeX | Étiquettes: ERIANI, Unité ARN
@article{,
title = {Fifteen years of the yeast three-hybrid system: RNA-protein interactions under investigation.},
author = {F Martin},
url = {http://www.ncbi.nlm.nih.gov/pubmed/22841566?dopt=Abstract},
doi = {10.1016/j.ymeth.2012.07.016},
isbn = {2284156},
year = {2012},
date = {2012-01-01},
journal = {Methods},
volume = {58},
number = {4},
pages = {367-375},
abstract = {In 1996, the Wickens and the Kuhl labs developed the yeast three-hybrid system independently. By expressing two chimeric proteins and one chimeric RNA molecule in Saccharomyces cerevisiae, this method allows in vivo monitoring of RNA-protein interactions by measuring the expression levels of HIS3 and LacZ reporter genes. Specific RNA targets have been used to characterize unknown RNA binding proteins. Previously described RNA binding proteins have also been used as bait to select new RNA targets. Finally, this method has been widely used to investigate or confirm previously suspected RNA-protein interactions. However, this method falls short in some aspects, such as RNA display and selection of false positive molecules. This review will summarize the results obtained with this method from the past 15years, as well as on recent efforts to improve its specificity.},
keywords = {ERIANI, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Huang Q, Yao P, Eriani G, Wang E D
In vivo identification of essential nucleotides in tRNALeu to its functions by using a constructed yeast tRNALeu knockout strain. Article de journal
Dans: Nucleic Acids Res, vol. 40, no. 20, p. 10463-10477, 2012, ISBN: 22917587.
Résumé | Liens | BibTeX | Étiquettes: ERIANI, Unité ARN
@article{,
title = {In vivo identification of essential nucleotides in tRNALeu to its functions by using a constructed yeast tRNALeu knockout strain.},
author = {Q Huang and P Yao and G Eriani and E D Wang},
url = {http://www.ncbi.nlm.nih.gov/pubmed/22917587?dopt=Abstract},
doi = {10.1093/nar/gks783},
isbn = {22917587},
year = {2012},
date = {2012-01-01},
journal = {Nucleic Acids Res},
volume = {40},
number = {20},
pages = {10463-10477},
abstract = {The fidelity of protein biosynthesis requires the aminoacylation of tRNA with its cognate amino acid catalyzed by aminoacyl-tRNA synthetase with high levels of accuracy and efficiency. Crucial bases in tRNA(Leu) to aminoacylation or editing functions of leucyl-tRNA synthetase have been extensively studied mainly by in vitro methods. In the present study, we constructed two Saccharomyces cerevisiae tRNA(Leu) knockout strains carrying deletions of the genes for tRNA(Leu)(GAG) and tRNA(Leu)(UAG). Disrupting the single gene encoding tRNA(Leu)(GAG) had no phenotypic consequence when compared to the wild-type strain. While disrupting the three genes for tRNA(Leu)(UAG) had a lethal effect on the yeast strain, indicating that tRNA(Leu)(UAG) decoding capacity could not be compensated by another tRNA(Leu) isoacceptor. Using the triple tRNA knockout strain and a randomly mutated library of tRNA(Leu)(UAG), a selection to identify critical tRNA(Leu) elements was performed. In this way, mutations inducing in vivo decreases of tRNA levels or aminoacylation or editing ability by leucyl-tRNA synthetase were identified. Overall, the data showed that the triple tRNA knockout strain is a suitable tool for in vivo studies and identification of essential nucleotides of the tRNA.},
keywords = {ERIANI, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Allmang C, Krol A
Seleneoprotein biosynthesis Chapitre d'ouvrage
Dans: Liu, J; Luo, G; Mu, Y (Ed.): Selenoproteins and Mimics, p. 107-124, Springer, 2012.
Résumé | Liens | BibTeX | Étiquettes: ERIANI, KROL, Unité ARN
@inbook{,
title = {Seleneoprotein biosynthesis},
author = {C Allmang and A Krol},
editor = {J Liu and G Luo and Y Mu},
url = {http://www.springerlink.com/content/q4h42729317k1478/},
doi = {10.1007/978-3-642-22236-8_8},
year = {2012},
date = {2012-01-01},
booktitle = {Selenoproteins and Mimics},
pages = {107-124},
publisher = {Springer},
series = {Advanced Topics and Technology in China},
abstract = {The amino acid selenocysteine (Sec) is the major biological form of the trace element selenium. Sec is co-translationally incorporated in selenoproteins and is found in the active site of those that have already been assigned a function. In eukaryotes, Sec biosynthesis from serine on the selenocysteine transfer RNA (tRNASec) requires four enzymes. The synthesis of selenoproteins follows a remarkable mechanism which involves translational recoding of a UGA codon, normally used as a stop signal, into a Sec codon. A surprisingly high number of molecular partners have been identified in this machinery but their mechanism of action is still largely unknown. In this chapter, we will provide a detailed description of the knowledge of the field.},
keywords = {ERIANI, KROL, Unité ARN},
pubstate = {published},
tppubtype = {inbook}
}
Zhou X L, Du D H, Tan M, Lei H Y, Ruan L L, Eriani G, Wang E D
Role of tRNA amino acid-accepting end in aminoacylation and its quality control Article de journal
Dans: Nucleic Acids Res, vol. 39, no. 20, p. 8857-8868, 2011, ISSN: 1362-4962 (Electronic) 0305-1048 (Linking), (DOI: 10.1093/nar/gkr595).
Résumé | Liens | BibTeX | Étiquettes: ERIANI, Unité ARN
@article{,
title = {Role of tRNA amino acid-accepting end in aminoacylation and its quality control},
author = {X L Zhou and D H Du and M Tan and H Y Lei and L L Ruan and G Eriani and E D Wang},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=21775341},
doi = {10.1093/nar/gkr595},
issn = {1362-4962 (Electronic)
0305-1048 (Linking)},
year = {2011},
date = {2011-01-01},
journal = {Nucleic Acids Res},
volume = {39},
number = {20},
pages = {8857-8868},
abstract = {Aminoacyl-tRNA synthetases (aaRSs) are remarkable enzymes that are in charge of the accurate recognition and ligation of amino acids and tRNA molecules. The greatest difficulty in accurate aminoacylation appears to be in discriminating between highly similar amino acids. To reduce mischarging of tRNAs by non-cognate amino acids, aaRSs have evolved an editing activity in a second active site to cleave the incorrect aminoacyl-tRNAs. Editing occurs after translocation of the aminoacyl-CCA(76) end to the editing site, switching between a hairpin and a helical conformation for aminoacylation and editing. Here, we studied the consequence of nucleotide changes in the CCA(76) accepting end of tRNA(Leu) during the aminoacylation and editing reactions. The analysis showed that the terminal A(76) is essential for both reactions, suggesting that critical interactions occur in the two catalytic sites. Substitutions of C(74) and C(75) selectively decreased aminoacylation keeping nearly unaffected editing. These mutations might favor the regular helical conformation required to reach the editing site. Mutating the editing domain residues that contribute to CCA(76) binding reduced the aminoacylation fidelity leading to cell-toxicity in the presence of non-cognate amino acids. Collectively, the data show how protein synthesis quality is controlled by the CCA(76) homogeneity of tRNAs.},
note = {DOI: 10.1093/nar/gkr595},
keywords = {ERIANI, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Zhu B, Yao P, Tan M, Eriani G, Wang E D
tRNA-independent Pretransfer Editing by Class I Leucyl-tRNA Synthetase Article de journal
Dans: J Biol Chem, vol. 284, no. 6, p. 3418-3424, 2009, ISBN: 19068478, (0021-9258 (Print) Journal Article).
Résumé | Liens | BibTeX | Étiquettes: ERIANI, Unité ARN
@article{,
title = {tRNA-independent Pretransfer Editing by Class I Leucyl-tRNA Synthetase},
author = {B Zhu and P Yao and M Tan and G Eriani and E D Wang},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19068478},
isbn = {19068478},
year = {2009},
date = {2009-01-01},
journal = {J Biol Chem},
volume = {284},
number = {6},
pages = {3418-3424},
abstract = {Aminoacyl-tRNA synthetases catalyze the formation of aminoacyl-tRNA in a two-step reaction starting with amino acid activation followed by aminoacyl group transfer to tRNA. To clear mistakes that occasionally occur, some of these enzymes carry out editing activities, acting on the misactivated amino acid (pretransfer editing) or after the transfer on the tRNA (post-transfer editing). The post-transfer editing pathway of leucyl-tRNA synthetase has been extensively studied by structural and biochemical approaches. Here, we report the finding of a tRNA-independent pretransfer editing pathway in leucyl-tRNA synthetases from Aquifex aeolicus. Using a CP1-mutant defective in its post-transfer editing function, we showed that this new editing pathway is distinct from the post-transfer editing site and may occur at the synthetic catalytic site, as recently proposed for other aminoacyl-tRNA synthetases.},
note = {0021-9258 (Print)
Journal Article},
keywords = {ERIANI, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Giege R, Eriani G
Transfer RNA recognition by synthetases Chapitre d'ouvrage
Dans: Encyclopedia of Life Sciences, John Wiley & Sons, 2009.
Résumé | Liens | BibTeX | Étiquettes: Aminoacyl-tRNA synthetase Genetic code Protein synthesis RNA recognition tRNA, ERIANI, Unité ARN
@inbook{,
title = {Transfer RNA recognition by synthetases},
author = {R Giege and G Eriani},
url = {http://www.els.net/WileyCDA/ElsArticle/refId-a0000531.html},
doi = {10.1002/9780470015902.a0000531.pub2},
year = {2009},
date = {2009-01-01},
booktitle = {Encyclopedia of Life Sciences},
publisher = {John Wiley & Sons},
abstract = {Fidelity of transfer ribonucleic acid (tRNA) charging by amino acids ensures correct translation of the genetic code into proteins. Charging is catalysed by a set of enzymes known as aminoacyl-tRNA synthetases. Owing to the degeneracy of the genetic code, some of the different tRNAs have the same amino acid attached to them. Specificity of the charging reaction is ensured by positive elements, the identity determinants unique to each tRNA and responsible for its recognition by the cognate synthetase, and negative elements, the antideterminants that prevent false recognitions. To fulfil the aminoacylation specificity and prevent noncognate aminoacyl-tRNA delivery to the ribosome, some synthetases also mediate proofreading reactions that increase fidelity of the tRNA charging. In such reactions, misactivated amino acids or mischarged tRNAs are checked in specific sites and noncognate products are hydrolysed.},
keywords = {Aminoacyl-tRNA synthetase Genetic code Protein synthesis RNA recognition tRNA, ERIANI, Unité ARN},
pubstate = {published},
tppubtype = {inbook}
}
Takeuchi A, Schmitt D, Chapple C, Babaylova E, Karpova G, Guigo R, Krol A, Allmang C
A short motif in Drosophila SECIS Binding Protein 2 provides differential binding affinity to SECIS RNA hairpins Article de journal
Dans: Nucleic Acids Res, vol. 37, no. 7, p. 2126-2141, 2009, ISBN: 19223320, (1362-4962 (Electronic) Journal Article Research Support, Non-U.S. Gov't).
Résumé | Liens | BibTeX | Étiquettes: ERIANI, KROL, Unité ARN
@article{,
title = {A short motif in Drosophila SECIS Binding Protein 2 provides differential binding affinity to SECIS RNA hairpins},
author = {A Takeuchi and D Schmitt and C Chapple and E Babaylova and G Karpova and R Guigo and A Krol and C Allmang},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19223320},
isbn = {19223320},
year = {2009},
date = {2009-01-01},
journal = {Nucleic Acids Res},
volume = {37},
number = {7},
pages = {2126-2141},
abstract = {Selenoproteins contain the amino acid selenocysteine which is encoded by a UGA Sec codon. Recoding UGA Sec requires a complex mechanism, comprising the cis-acting SECIS RNA hairpin in the 3'UTR of selenoprotein mRNAs, and trans-acting factors. Among these, the SECIS Binding Protein 2 (SBP2) is central to the mechanism. SBP2 has been so far functionally characterized only in rats and humans. In this work, we report the characterization of the Drosophila melanogaster SBP2 (dSBP2). Despite its shorter length, it retained the same selenoprotein synthesis-promoting capabilities as the mammalian counterpart. However, a major difference resides in the SECIS recognition pattern: while human SBP2 (hSBP2) binds the distinct form 1 and 2 SECIS RNAs with similar affinities, dSBP2 exhibits high affinity toward form 2 only. In addition, we report the identification of a K (lysine)-rich domain in all SBP2s, essential for SECIS and 60S ribosomal subunit binding, differing from the well-characterized L7Ae RNA-binding domain. Swapping only five amino acids between dSBP2 and hSBP2 in the K-rich domain conferred reversed SECIS-binding properties to the proteins, thus unveiling an important sequence for form 1 binding.},
note = {1362-4962 (Electronic)
Journal Article
Research Support, Non-U.S. Gov't},
keywords = {ERIANI, KROL, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Olieric V, Wolff P, Takeuchi A, Bec G, Birck C, Vitorino M, Kieffer B, Beniaminov A, Cavigiolio G, Theil E, Allmang C, Krol A, Dumas P
SECIS-binding protein 2, a key player in selenoprotein synthesis, is an intrinsically disordered protein Article de journal
Dans: Biochimie, vol. 91, no. 8, p. 1003-1009, 2009, ISBN: 19467292, (1638-6183 (Electronic) Journal Article Research Support, Non-U.S. Gov't).
Résumé | Liens | BibTeX | Étiquettes: ENNIFAR, ERIANI, Unité ARN
@article{,
title = {SECIS-binding protein 2, a key player in selenoprotein synthesis, is an intrinsically disordered protein},
author = {V Olieric and P Wolff and A Takeuchi and G Bec and C Birck and M Vitorino and B Kieffer and A Beniaminov and G Cavigiolio and E Theil and C Allmang and A Krol and P Dumas},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19467292},
isbn = {19467292},
year = {2009},
date = {2009-01-01},
journal = {Biochimie},
volume = {91},
number = {8},
pages = {1003-1009},
abstract = {Selenocysteine (Sec) is co-translationally incorporated into selenoproteins at a reprogrammed UGA codon. In mammals, this requires a dedicated machinery comprising a stem-loop structure in the 3' UTR RNA (the SECIS element) and the specific SECIS Binding Protein 2. In this report, disorder-prediction methods and several biophysical techniques showed that ca. 70% of the SBP2 sequence is disordered, whereas the RNA binding domain appears to be folded and functional. These results are consistent with a recent report on the role of the Hsp90 chaperone for the folding of SBP2 and other functionally unrelated proteins bearing an RNA binding domain homologous to SBP2.},
note = {1638-6183 (Electronic)
Journal Article
Research Support, Non-U.S. Gov't},
keywords = {ENNIFAR, ERIANI, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Allmang C, Wurth L, Krol A
The selenium to selenoprotein pathway in eukaryotes: more molecular partners than anticipated Article de journal
Dans: Biochim Biophys Acta-Gen Subj, vol. 1790, no. 11, p. 1415-1423, 2009, ISBN: 19285539, (0006-3002 (Print) 0006-3002 (Linking) Journal Article Review).
Résumé | Liens | BibTeX | Étiquettes: Animals Base Sequence Eukaryota/genetics/*metabolism Humans Metabolic Networks and Pathways/genetics/physiology Models, Biological Selenium/*metabolism Selenocysteine/biosynthesis Selenoproteins/biosynthesis/genetics/*metabolism, ERIANI, Unité ARN
@article{,
title = {The selenium to selenoprotein pathway in eukaryotes: more molecular partners than anticipated},
author = {C Allmang and L Wurth and A Krol},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19285539},
isbn = {19285539},
year = {2009},
date = {2009-01-01},
journal = {Biochim Biophys Acta-Gen Subj},
volume = {1790},
number = {11},
pages = {1415-1423},
abstract = {The amino acid selenocysteine (Sec) is the major biological form of the trace element selenium. Sec is co-translationally incorporated in selenoproteins. There are 25 selenoprotein genes in humans, and Sec was found in the active site of those that have been attributed a function. This review will discuss how selenocysteine is synthesized and incorporated into selenoproteins in eukaryotes. Sec biosynthesis from serine on the tRNA(Sec) requires four enzymes. Incorporation of Sec in response to an in-frame UGA codon, otherwise signaling termination of translation, is achieved by a complex recoding machinery to inform the ribosomes not to stop at this position on the mRNA. A number of the molecular partners acting in this machinery have been identified but their detailed mechanism of action has not been deciphered yet. Here we provide an overview of the literature in the field. Particularly striking is the higher than originally envisaged number of factors necessary to synthesize Sec and selenoproteins. Clearly, selenoprotein synthesis is an exciting and very active field of research.},
note = {0006-3002 (Print)
0006-3002 (Linking)
Journal Article
Review},
keywords = {Animals Base Sequence Eukaryota/genetics/*metabolism Humans Metabolic Networks and Pathways/genetics/physiology Models, Biological Selenium/*metabolism Selenocysteine/biosynthesis Selenoproteins/biosynthesis/genetics/*metabolism, ERIANI, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Yao P, Zhu B, Jaeger S, Eriani G, Wang E D
Recognition of tRNALeu by Aquifex aeolicus leucyl-tRNA synthetase during the aminoacylation and editing steps Article de journal
Dans: Nucleic Acids Res, vol. 36, no. 8, p. 2728-2738, 2008, ISBN: 18367476, (1362-4962 (Electronic) Journal Article Research Support, Non-U.S. Gov't).
Résumé | Liens | BibTeX | Étiquettes: Amino Acids/chemistry Anticodon/chemistry Bacteria/enzymology/genetics Base Sequence Iodine Leucine-tRNA Ligase/chemistry/*metabolism Molecular Sequence Data Mutagenesis Nucleic Acid Conformation Protein Footprinting RNA, ERIANI, Leu/*chemistry/genetics/metabolism Ribonucleases Substrate Specificity *Transfer RNA Aminoacylation, Transfer, Unité ARN
@article{,
title = {Recognition of tRNALeu by Aquifex aeolicus leucyl-tRNA synthetase during the aminoacylation and editing steps},
author = {P Yao and B Zhu and S Jaeger and G Eriani and E D Wang},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18367476},
isbn = {18367476},
year = {2008},
date = {2008-01-01},
journal = {Nucleic Acids Res},
volume = {36},
number = {8},
pages = {2728-2738},
abstract = {Recognition of tRNA by the cognate aminoacyl-tRNA synthetase during translation is crucial to ensure the correct expression of the genetic code. To understand tRNA(Leu) recognition sets and their evolution, the recognition of tRNA(Leu) by the leucyl-tRNA synthetase (LeuRS) from the primitive hyperthermophilic bacterium Aquifex aeolicus was studied by RNA probing and mutagenesis. The results show that the base A73; the core structure of tRNA formed by the tertiary interactions U8-A14, G18-U55 and G19-C56; and the orientation of the variable arm are critical elements for tRNA(Leu) aminoacylation. Although dispensable for aminoacylation, the anticodon arm carries discrete editing determinants that are required for stabilizing the conformation of the post-transfer editing state and for promoting translocation of the tRNA acceptor arm from the synthetic to the editing site.},
note = {1362-4962 (Electronic)
Journal Article
Research Support, Non-U.S. Gov't},
keywords = {Amino Acids/chemistry Anticodon/chemistry Bacteria/enzymology/genetics Base Sequence Iodine Leucine-tRNA Ligase/chemistry/*metabolism Molecular Sequence Data Mutagenesis Nucleic Acid Conformation Protein Footprinting RNA, ERIANI, Leu/*chemistry/genetics/metabolism Ribonucleases Substrate Specificity *Transfer RNA Aminoacylation, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Boulon S, Marmier-Gourrier N, Pradet-Balade B, Wurth L, Verheggen C, Jady B E, Rothé B, Pescia C, Robert M C, Kiss T, Bardoni B, Krol A, Branlant C, Allmang C, Bertrand E, Charpentier B
The Hsp90 chaperone controls the biogenesis of L7Ae RNPs through conserved machinery Article de journal
Dans: J Cell Biol, vol. 180, no. 3, p. 579-595, 2008, ISSN: 0021-9525, (1540-8140 (Electronic) Journal Article Research Support, Non-U.S. Gov't).
Résumé | Liens | BibTeX | Étiquettes: ERIANI, KROL Adenosine Triphosphatases/genetics/metabolism Cell Cycle Proteins/genetics/metabolism Cell Proliferation Conserved Sequence/genetics DNA Helicases/genetics/metabolism DNA-Binding Proteins/genetics/metabolism Evolution, Unité ARN
@article{,
title = {The Hsp90 chaperone controls the biogenesis of L7Ae RNPs through conserved machinery},
author = {S Boulon and N Marmier-Gourrier and B Pradet-Balade and L Wurth and C Verheggen and B E Jady and B Rothé and C Pescia and M C Robert and T Kiss and B Bardoni and A Krol and C Branlant and C Allmang and E Bertrand and B Charpentier},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18268104},
issn = {0021-9525},
year = {2008},
date = {2008-01-01},
journal = {J Cell Biol},
volume = {180},
number = {3},
pages = {579-595},
abstract = {RNA-binding proteins of the L7Ae family are at the heart of many essential ribonucleoproteins (RNPs), including box C/D and H/ACA small nucleolar RNPs, U4 small nuclear RNP, telomerase, and messenger RNPs coding for selenoproteins. In this study, we show that Nufip and its yeast homologue Rsa1 are key components of the machinery that assembles these RNPs. We observed that Rsa1 and Nufip bind several L7Ae proteins and tether them to other core proteins in the immature particles. Surprisingly, Rsa1 and Nufip also link assembling RNPs with the AAA + adenosine triphosphatases hRvb1 and hRvb2 and with the Hsp90 chaperone through two conserved adaptors, Tah1/hSpagh and Pih1. Inhibition of Hsp90 in human cells prevents the accumulation of U3, U4, and telomerase RNAs and decreases the levels of newly synthesized hNop58, hNHP2, 15.5K, and SBP2. Thus, Hsp90 may control the folding of these proteins during the formation of new RNPs. This suggests that Hsp90 functions as a master regulator of cell proliferation by allowing simultaneous control of cell signaling and cell growth.},
note = {1540-8140 (Electronic)
Journal Article
Research Support, Non-U.S. Gov't},
keywords = {ERIANI, KROL Adenosine Triphosphatases/genetics/metabolism Cell Cycle Proteins/genetics/metabolism Cell Proliferation Conserved Sequence/genetics DNA Helicases/genetics/metabolism DNA-Binding Proteins/genetics/metabolism Evolution, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Zhu B, Zhao M W, Eriani G, Wang E D
A present-day aminoacyl-tRNA synthetase with ancestral editing properties Article de journal
Dans: RNA, vol. 13, no. 1, p. 15-21, 2007, ISBN: 17095543, (1355-8382 (Print) Journal Article Research Support, Non-U.S. Gov't).
Résumé | Liens | BibTeX | Étiquettes: ERIANI, Unité ARN
@article{,
title = {A present-day aminoacyl-tRNA synthetase with ancestral editing properties},
author = {B Zhu and M W Zhao and G Eriani and E D Wang},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17095543},
isbn = {17095543},
year = {2007},
date = {2007-01-01},
journal = {RNA},
volume = {13},
number = {1},
pages = {15-21},
abstract = {Leucyl-, isoleucyl-, and valyl-tRNA synthetases form a subgroup of related aminoacyl-tRNA synthetases that attach similar amino acids to their cognate tRNAs. To prevent amino acid misincorporation during translation, these enzymes also hydrolyze mischarged tRNAs through a post-transfer editing mechanism. Here we show that LeuRS from the deep-branching bacterium Aquifex aeolicus edits the complete set of aminoacylated tRNAs generated by the three enzymes: Ile-tRNA(Ile), Val-tRNA(Ile), Val-tRNA(Val), Thr-tRNA(Val), and Ile-tRNA(Leu). This unusual enlarged editing property was studied in a model of a primitive editing system containing a composite minihelix carrying the triple leucine, isoleucine, and valine identity mimicking the primitive tRNA precursor. We found that the freestanding LeuRS editing domain can edit this precursor in contrast to IleRS and ValRS editing domains. These results suggest that A. aeolicus LeuRS carries editing properties that seem more primitive than those of IleRS and ValRS. They suggest that the A. aeolicus editing domain has preserved the ambiguous editing property from the ancestral common editing domain or, alternatively, that this plasticity results from a specific metabolic adaptation.},
note = {1355-8382 (Print)
Journal Article
Research Support, Non-U.S. Gov't},
keywords = {ERIANI, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Olieric N, Bey G, Nierengarten H, Wang E D, Moras D, Eriani G, Cavarelli J
Expression, purification, and characterization of a new heterotetramer structure of leucyl-tRNA synthetase from Aquifex aeolicus in Escherichia coli Article de journal
Dans: Protein Expr Purif, vol. 47, no. 1, p. 1-9, 2006, ISBN: 16256368, (1046-5928 (Print) Journal Article).
Résumé | Liens | BibTeX | Étiquettes: ERIANI, Unité ARN
@article{,
title = {Expression, purification, and characterization of a new heterotetramer structure of leucyl-tRNA synthetase from Aquifex aeolicus in Escherichia coli},
author = {N Olieric and G Bey and H Nierengarten and E D Wang and D Moras and G Eriani and J Cavarelli},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16256368},
isbn = {16256368},
year = {2006},
date = {2006-01-01},
journal = {Protein Expr Purif},
volume = {47},
number = {1},
pages = {1-9},
abstract = {Aminoacyl-tRNA synthetases are key players in the interpretation of the genetic code. They constitute a textbook example of multi-domain proteins including insertion and terminal functional modules appended to one of the two class-specific active site domains. The non-catalytic domains usually have distinct roles in the aminoacylation reaction. Aquifex aeolicus leucyl-tRNA synthetase (LeuRS) is composed of a separated catalytic site and tRNA anticodon-binding site, which would represent one of the closest relics of the primordial aminoacyl-tRNA synthetase. Moreover, the essential catalytic site residues are split into the two different subunits. In all other class-I aminoacyl-tRNA synthetases, those two functional polypeptides are nowadays fused into a single protein chain. In this work, we report the isolation and the characterization, in Escherichia coli, of a novel oligomeric form (alphabeta)2 for A. aeolicus LeuRS, which is present in addition to the alphabeta heterodimer. A. aeolicus (alphabeta)2 LeuRS has been characterized by biochemical and biophysical methods. Native gel electrophoresis, mass spectrometry, analytical ultracentrifugation, and kinetic analysis confirmed that the (alphabeta)2 enzyme was a stable and active entity. By mass spectrometry we confirmed that the heterodimer alphabeta can bind one tRNALeu molecule whereas the heterotetramer (alphabeta)2 can bind two tRNALeu molecules. Active site titration and aminoacylation assays showed that two functional active sites are found per heterotetramer, suggesting that this molecular species might exist and be active in vivo. All those data suggest that the existence of the heterotetramer is certainly not an artifact of overexpression in E. coli.},
note = {1046-5928 (Print)
Journal Article},
keywords = {ERIANI, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Jaeger S, Martin F, Rudinger-Thirion J, Giege R, Eriani G
Binding of human SLBP on the 3'-UTR of histone precursor H4-12 mRNA induces structural rearrangements that enable U7 snRNA anchoring Article de journal
Dans: Nucleic Acids Res, vol. 34, no. 17, p. 4987-4995, 2006, ISBN: 16982637, (1362-4962 (Electronic) Journal Article).
Résumé | Liens | BibTeX | Étiquettes: ERIANI, FRUGIER, Unité ARN
@article{,
title = {Binding of human SLBP on the 3'-UTR of histone precursor H4-12 mRNA induces structural rearrangements that enable U7 snRNA anchoring},
author = {S Jaeger and F Martin and J Rudinger-Thirion and R Giege and G Eriani},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16982637},
isbn = {16982637},
year = {2006},
date = {2006-01-01},
journal = {Nucleic Acids Res},
volume = {34},
number = {17},
pages = {4987-4995},
abstract = {In metazoans, cell-cycle-dependent histones are produced from poly(A)-lacking mRNAs. The 3' end of histone mRNAs is formed by an endonucleolytic cleavage of longer precursors between a conserved stem-loop structure and a purine-rich histone downstream element (HDE). The cleavage requires at least two trans-acting factors: the stem-loop binding protein (SLBP), which binds to the stem-loop and the U7 snRNP, which anchors to histone pre-mRNAs by annealing to the HDE. Using RNA structure-probing techniques, we determined the secondary structure of the 3'-untranslated region (3'-UTR) of mouse histone pre-mRNAs H4-12, H1t and H2a-614. Surprisingly, the HDE is embedded in hairpin structures and is therefore not easily accessible for U7 snRNP anchoring. Probing of the 3'-UTR in complex with SLBP revealed structural rearrangements leading to an overall opening of the structure especially at the level of the HDE. Electrophoretic mobility shift assays demonstrated that the SLBP-induced opening of HDE actually facilitates U7 snRNA anchoring on the histone H4-12 pre-mRNAs 3' end. These results suggest that initial binding of the SLBP functions in making the HDE more accessible for U7 snRNA anchoring.},
note = {1362-4962 (Electronic)
Journal Article},
keywords = {ERIANI, FRUGIER, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Zhao M W, Zhu B, Hao R, Xu M G, Eriani G, Wang E D
Leucyl-tRNA synthetase from the ancestral bacterium Aquifex aeolicus contains relics of synthetase evolution Article de journal
Dans: EMBO J, vol. 24, no. 7, p. 1430-1439, 2005, ISBN: 15775966, (0261-4189 (Print) Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Amino Acid Sequence Bacteria/*genetics Comparative Study Escherichia coli/genetics *Evolution, ERIANI, Molecular Leucine-tRNA Ligase/*genetics/*metabolism Models, Molecular Molecular Sequence Data Protein Biosynthesis/*genetics Protein Structure, Non-U.S. Gov't Sequence Alignment, Tertiary/genetics Research Support, Unité ARN
@article{,
title = {Leucyl-tRNA synthetase from the ancestral bacterium Aquifex aeolicus contains relics of synthetase evolution},
author = {M W Zhao and B Zhu and R Hao and M G Xu and G Eriani and E D Wang},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15775966},
isbn = {15775966},
year = {2005},
date = {2005-01-01},
journal = {EMBO J},
volume = {24},
number = {7},
pages = {1430-1439},
abstract = {The editing reactions catalyzed by aminoacyl-tRNA synthetases are critical for the faithful protein synthesis by correcting misactivated amino acids and misaminoacylated tRNAs. We report that the isolated editing domain of leucyl-tRNA synthetase from the deep-rooted bacterium Aquifex aeolicus (alphabeta-LeuRS) catalyzes the hydrolytic editing of both mischarged tRNA(Leu) and minihelix(Leu). Within the domain, we have identified a crucial 20-amino-acid peptide that confers editing capacity when transplanted into the inactive Escherichia coli LeuRS editing domain. Likewise, fusion of the beta-subunit of alphabeta-LeuRS to the E. coli editing domain activates its editing function. These results suggest that alphabeta-LeuRS still carries the basic features from a primitive synthetase molecule. It has a remarkable capacity to transfer autonomous active modules, which is consistent with the idea that modern synthetases arose after exchange of small idiosyncratic domains. It also has a unique alphabeta-heterodimeric structure with separated catalytic and tRNA-binding sites. Such an organization supports the tRNA/synthetase coevolution theory that predicts sequential addition of tRNA and synthetase domains.},
note = {0261-4189 (Print)
Journal Article},
keywords = {Amino Acid Sequence Bacteria/*genetics Comparative Study Escherichia coli/genetics *Evolution, ERIANI, Molecular Leucine-tRNA Ligase/*genetics/*metabolism Models, Molecular Molecular Sequence Data Protein Biosynthesis/*genetics Protein Structure, Non-U.S. Gov't Sequence Alignment, Tertiary/genetics Research Support, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Jaeger S, Barends S, Giege R, Eriani G, Martin F
Expression of metazoan replication-dependent histone genes Article de journal
Dans: Biochimie, vol. 87, no. 9-10, p. 827-834, 2005, ISBN: 16164992, (0300-9084 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: ERIANI, Unité ARN
@article{,
title = {Expression of metazoan replication-dependent histone genes},
author = {S Jaeger and S Barends and R Giege and G Eriani and F Martin},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16164992},
isbn = {16164992},
year = {2005},
date = {2005-01-01},
journal = {Biochimie},
volume = {87},
number = {9-10},
pages = {827-834},
abstract = {Histone proteins are essential components of eukaryotic chromosomes. In metazoans, they are produced from the so-called replication-dependent histone genes. The biogenesis of histones is tightly coupled to DNA replication in a stoichiometric manner because an excess of histones is highly toxic for the cell. Therefore, a strict cell cycle-regulation of critical factors required for histone expression ensures exclusive S-phase expression. This review focuses on the molecular mechanisms responsible for such a fine expression regulation. Among these, a large part will be dedicated to post-transcriptional events occurring on histone mRNA, like histone mRNA 3' end processing, nucleo-cytoplasmic mRNA export, translation and mRNA degradation. Many factors are involved, including an RNA-binding protein called HBP, also called SLBP (for hairpin- or stem-loop-binding protein) that binds to a conserved hairpin located in the 3' UTR part of histone mRNA. HBP plays a pivotal role in the expression of histone genes since it is necessary for most of the steps of histone mRNA metabolism in the cell. Moreover, the strict S-phase expression pattern of histones is achieved through a fine cell cycle-regulation of HBP. A large part of the discussion will be centered on the critical role of HBP in histone biogenesis.},
note = {0300-9084
Journal Article},
keywords = {ERIANI, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Eriani G, Cavarelli J
Arginyl-tRNA synthetase Chapitre d'ouvrage
Dans: Ibba, M; Francklyn, C; Cusack, S (Ed.): The Aminoacyl-tRNA Synthetases, Landes Bioscience, 2005.
Résumé | Liens | BibTeX | Étiquettes: ERIANI, Unité ARN
@inbook{,
title = {Arginyl-tRNA synthetase},
author = {G Eriani and J Cavarelli},
editor = {M Ibba and C Francklyn and S Cusack},
url = {http://www.landesbioscience.com/curie/chapter/1284},
year = {2005},
date = {2005-01-01},
booktitle = {The Aminoacyl-tRNA Synthetases},
publisher = {Landes Bioscience},
abstract = {Determination of the crystal structures of arginyl-tRNA synthetase, either in the free state or engaged in complexes with the other partners of the arginylation reaction, led to fundamental progress in understanding the sequence-structure-function relationship of this catalytic reaction. The structures reveal unexpected results simplifying and organizing the collected biological information but also illustrating the inherent complexity of this macromolecular recognition process. ArgRS specifically recognizes the D-loop and the anticodon of tRNAArg using dedicated modules. Conformational changes which occur upon substrates binding have been visualized at the atomic level. While ArgRS requires its cognate tRNA for the first step of the aminoacylation reaction, the crystal structures reveal that (i) L-arginine binding controls the correct positioning of the CCA end of tRNAArg and that (ii) tRNAArg binding produces conformational changes of the ATP-binding cleft. In this review, results from extensive investigations preformed by several groups are summarized.},
keywords = {ERIANI, Unité ARN},
pubstate = {published},
tppubtype = {inbook}
}
Cura V, Olieric N, Guichard A, Wang E D, Moras D, Eriani G, Cavarelli J
Crystallization and preliminary X-ray crystallographic study of the wild type and two mutants of the CP1 hydrolytic domain from Aquifex aeolicus leucyl-tRNA synthetase Article de journal
Dans: Acta Crystallogr F Struct Biol Commun, vol. 61, no. Pt 10, p. 899-901, 2005, ISBN: 16511190, (1744-3091 (Electronic) Journal Article).
Résumé | Liens | BibTeX | Étiquettes: ERIANI, Unité ARN
@article{,
title = {Crystallization and preliminary X-ray crystallographic study of the wild type and two mutants of the CP1 hydrolytic domain from Aquifex aeolicus leucyl-tRNA synthetase},
author = {V Cura and N Olieric and A Guichard and E D Wang and D Moras and G Eriani and J Cavarelli},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16511190},
isbn = {16511190},
year = {2005},
date = {2005-01-01},
journal = {Acta Crystallogr F Struct Biol Commun},
volume = {61},
number = {Pt 10},
pages = {899-901},
abstract = {The editing or hydrolytic CP1 domain of leucyl-tRNA synthetase (LeuRS) hydrolyses several misactivated amino acids. The CP1 domain of Aquifex aeolicus LeuRS was expressed, purified and crystallized by the hanging-drop vapour-diffusion method using ammonium sulfate as precipitant. Crystals belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 38.8},
note = {1744-3091 (Electronic)
Journal Article},
keywords = {ERIANI, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Sissler M, Helm M, Frugier M, Giege R, Florentz C
Aminoacylation properties of pathology-related human mitochondrial tRNA(Lys) variants Article de journal
Dans: RNA, vol. 10, no. 5, p. 841-853, 2004, ISBN: 15100439, (1355-8382 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: ERIANI, FLORENTZ, FLORENTZ GIEGE Acylation Aminoacyltransferases/*genetics Human MERRF Syndrome/genetics Mitochondria/*genetics Mitochondrial Diseases/*genetics Mutation Nucleic Acid Conformation RNA, FRUGIER, Lys/*genetics Sequence Analysis, Non-U.S. Gov't Variation (Genetics), RNA Support, SISSLER, Transfer, Unité ARN
@article{,
title = {Aminoacylation properties of pathology-related human mitochondrial tRNA(Lys) variants},
author = {M Sissler and M Helm and M Frugier and R Giege and C Florentz},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15100439},
isbn = {15100439},
year = {2004},
date = {2004-01-01},
journal = {RNA},
volume = {10},
number = {5},
pages = {841-853},
abstract = {In vitro transcription has proven to be a successful tool for preparation of functional RNAs, especially in the tRNA field, in which, despite the absence of post-transcriptional modifications, transcripts are correctly folded and functionally active. Human mitochondrial (mt) tRNA(Lys) deviates from this principle and folds into various inactive conformations, due to the absence of the post-transcriptional modification m(1)A9 which hinders base-pairing with U64 in the native tRNA. Unavailability of a functional transcript is a serious drawback for structure/function investigations as well as in deciphering the molecular mechanisms by which point mutations in the mt tRNA(Lys) gene cause severe human disorders. Here, we show that an engineered in vitro transcribed "pseudo-WT" tRNA(Lys) variant is efficiently recognized by lysyl-tRNA synthetase and can substitute for the WT tRNA as a valuable reference molecule. This has been exploited in a systematic analysis of the effects on aminoacylation of nine pathology-related mutations described so far. The sole mutation located in a loop of the tRNA secondary structure, A8344G, does not affect aminoacylation efficiency. Out of eight mutations located in helical domains converting canonical Watson-Crick pairs into G-U pairs or C.A mismatches, six have no effect on aminoacylation (A8296G, U8316C, G8342A, U8356C, U8362G, G8363A), and two lead to drastic decreases (5000- to 7000-fold) in lysylation efficiencies (G8313A and G8328A). This screening, allowing for analysis of the primary impact level of all mutations affecting one tRNA under comparable conditions, indicates distinct molecular origins for different disorders.},
note = {1355-8382
Journal Article},
keywords = {ERIANI, FLORENTZ, FLORENTZ GIEGE Acylation Aminoacyltransferases/*genetics Human MERRF Syndrome/genetics Mitochondria/*genetics Mitochondrial Diseases/*genetics Mutation Nucleic Acid Conformation RNA, FRUGIER, Lys/*genetics Sequence Analysis, Non-U.S. Gov't Variation (Genetics), RNA Support, SISSLER, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Zheng Y G, Wei H, Ling C, Martin F, Eriani G, Wang E D
Two distinct domains of the beta subunit of Aquifex aeolicus leucyl-tRNA synthetase are involved in tRNA binding as revealed by a three-hybrid selection Article de journal
Dans: Nucleic Acids Res, vol. 32, no. 11, p. 3294-3303, 2004, ISBN: 15208367, (1362-4962 Evaluation Studies Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Amino Acid Sequence Binding Sites Gram-Negative Bacteria/*enzymology Kinetics Leucine-tRNA Ligase/*chemistry/genetics/*metabolism Molecular Sequence Data Mutation Protein Structure, ERIANI, Leu/*metabolism Saccharomyces cerevisiae/genetics *Two-Hybrid System Techniques, Tertiary Protein Subunits/chemistry RNA, Transfer, Unité ARN
@article{,
title = {Two distinct domains of the beta subunit of Aquifex aeolicus leucyl-tRNA synthetase are involved in tRNA binding as revealed by a three-hybrid selection},
author = {Y G Zheng and H Wei and C Ling and F Martin and G Eriani and E D Wang},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15208367},
isbn = {15208367},
year = {2004},
date = {2004-01-01},
journal = {Nucleic Acids Res},
volume = {32},
number = {11},
pages = {3294-3303},
abstract = {The Aquifex aeolicus alphabeta-LeuRS is the only known heterodimeric class Ia aminoacyl-tRNA synthetase. In this study, we investigated the function of the beta subunit which is believed to bind tRNA(Leu). A yeast three-hybrid system was constructed on the basis of the interaction of the beta subunit with its cognate tRNA(Leu). Then, seven mutated beta subunits exhibiting impaired tRNA binding capacities were selected out from a randomly mutated library. Two mutations were identified in the class Ia-helix-bundle-domain, which might interact with the D-hairpin of the tRNA analogous to other class Ia tRNA:synthetases complexes. The five other mutations were found in the LeuRS-specific C-terminal domain of which the folding is still unknown. tRNA affinity measurements and kinetic analyses performed on the isolated beta subunits and on the co-expressed alphabeta-heterodimers showed for all the mutants an effect in tRNA affinity in the ground state. In addition, an effect on the transition state of the aminoacylation reaction was observed for a 21-residues deletion mutant of the C-terminal end. These results show that the genetic approach of the three hybrid system is widely applicable and is a powerful tool for the investigation of tRNA:synthetase interactions.},
note = {1362-4962
Evaluation Studies
Journal Article},
keywords = {Amino Acid Sequence Binding Sites Gram-Negative Bacteria/*enzymology Kinetics Leucine-tRNA Ligase/*chemistry/genetics/*metabolism Molecular Sequence Data Mutation Protein Structure, ERIANI, Leu/*metabolism Saccharomyces cerevisiae/genetics *Two-Hybrid System Techniques, Tertiary Protein Subunits/chemistry RNA, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Martin F, Barends S, Eriani G
Single amino acid changes in AspRS reveal alternative routes for expanding its tRNA repertoire in vivo Article de journal
Dans: Nucleic Acids Res, vol. 32, no. 13, p. 4081-4089, 2004, ISBN: 15289581, (1362-4962 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Amino Acid Substitution Anticodon/metabolism Aspartate-tRNA Ligase/*chemistry/genetics/*metabolism Aspartic Acid/metabolism Binding Sites Models, Amino Acyl/chemistry/*metabolism Support, ERIANI, Molecular Mutation Phenotype Protein Engineering Protein Structure, Non-U.S. Gov't, Tertiary RNA, Transfer, Unité ARN
@article{,
title = {Single amino acid changes in AspRS reveal alternative routes for expanding its tRNA repertoire in vivo},
author = {F Martin and S Barends and G Eriani},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15289581},
isbn = {15289581},
year = {2004},
date = {2004-01-01},
journal = {Nucleic Acids Res},
volume = {32},
number = {13},
pages = {4081-4089},
abstract = {Aminoacyl-tRNA synthetases (aaRSs) are enzymes that are highly specific for their tRNA substrates. Here, we describe the expansion of a class IIb aaRS-tRNA specificity by a genetic selection that involves the use of a modified tRNA displaying an amber anticodon and the argE(amber) and lacZ(amber) reporters. The study was performed on Escherichia coli aspartyl-tRNA synthetase (AspRS) and amber tRNA(Asp). Nine AspRS mutants able to charge the amber tRNA(Asp) and to suppress the reporter genes were selected from a randomly mutated library. All the mutants exhibited a new amber tRNA(Asp) specificity in addition to the initial native tRNA(Asp). Six mutations were found in the anticodon-binding site located in the N-terminal OB-fold. The strongest suppressor was a mutation of residue Glu-93 that contacts specifically the anticodon nucleotide 34 in the crystal structure. The other mutations in the OB-fold were found at close distance from the anticodon in the so-called loop L45 and strand S1. They concern residues that do not contact tRNA(Asp) in the native complex. In addition, this study shows that suppressors can carry mutations located far from the anticodon-binding site. One such mutation was found in the synthetase hinge-module where it increases the tRNA(Asp)-charging rate, and two other mutations were found in the prokaryotic-specific insertion domain and the catalytic core. These mutants seem to act by indirect effects on the tRNA acceptor stem binding and on the conformation of the active site of the enzyme. Altogether, these data suggest the existence of various ways for modifying the mechanism of tRNA discrimination.},
note = {1362-4962
Journal Article},
keywords = {Amino Acid Substitution Anticodon/metabolism Aspartate-tRNA Ligase/*chemistry/genetics/*metabolism Aspartic Acid/metabolism Binding Sites Models, Amino Acyl/chemistry/*metabolism Support, ERIANI, Molecular Mutation Phenotype Protein Engineering Protein Structure, Non-U.S. Gov't, Tertiary RNA, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Jaeger S, Eriani G, Martin F
Results and prospects of the yeast three-hybrid system Article de journal
Dans: FEBS Lett, vol. 556, no. 1-3, p. 7-12, 2004, ISBN: 14706817, (0014-5793 Journal Article Review Review Literature).
Résumé | Liens | BibTeX | Étiquettes: ERIANI, Histones/genetics Human RNA/chemistry/genetics/metabolism RNA-Binding Proteins/chemistry/genetics/metabolism Saccharomyces cerevisiae/*genetics Support, Non-U.S. Gov't *Two-Hybrid System Techniques, Unité ARN
@article{,
title = {Results and prospects of the yeast three-hybrid system},
author = {S Jaeger and G Eriani and F Martin},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14706817},
isbn = {14706817},
year = {2004},
date = {2004-01-01},
journal = {FEBS Lett},
volume = {556},
number = {1-3},
pages = {7-12},
abstract = {In 1996, a new method, termed the yeast three-hybrid system, dedicated to selection of RNA binding proteins using a hybrid RNA molecule as bait was described. In this minireview, we summarize the results that have been obtained using this method. Indeed, approximately 20 unknown proteins have been characterized so far. The three-hybrid strategy has also been used as a tool to dissect RNA-protein interactions. The example of such a study on human histone HBP interaction with its target mRNA is described. Problems that can be encountered are addressed in a troubleshooting section. Especially, our results with tRNA binding proteins are discussed.},
note = {0014-5793
Journal Article
Review
Review Literature},
keywords = {ERIANI, Histones/genetics Human RNA/chemistry/genetics/metabolism RNA-Binding Proteins/chemistry/genetics/metabolism Saccharomyces cerevisiae/*genetics Support, Non-U.S. Gov't *Two-Hybrid System Techniques, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Jaeger S, Eriani G, Martin F
Critical residues for RNA discrimination of the histone hairpin binding protein (HBP) investigated by the yeast three-hybrid system Article de journal
Dans: FEBS Lett, vol. 556, no. 1-3, p. 265-270, 2004, ISBN: 14706861, (0014-5793 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Amino Acid Support, Amino Acid Sequence Amino Acid Substitution Base Sequence Carrier Proteins/chemistry/*genetics/*metabolism Histones/genetics Human Lac Operon/genetics Molecular Sequence Data Nucleic Acid Conformation Nucleotides/chemistry/genetics/metabolism RNA/chemistry/genetics/*metabolism RNA, ERIANI, Messenger/chemistry/genetics/metabolism RNA-Binding Proteins/chemistry/genetics/metabolism Recombinant Fusion Proteins/chemistry/genetics/metabolism Saccharomyces cerevisiae/*genetics Sequence Alignment Sequence Homology, Non-U.S. Gov't Two-Hybrid System Techniques beta-Galactosidase/genetics/metabolism, Unité ARN
@article{,
title = {Critical residues for RNA discrimination of the histone hairpin binding protein (HBP) investigated by the yeast three-hybrid system},
author = {S Jaeger and G Eriani and F Martin},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14706861},
isbn = {14706861},
year = {2004},
date = {2004-01-01},
journal = {FEBS Lett},
volume = {556},
number = {1-3},
pages = {265-270},
abstract = {The histone hairpin binding protein (HBP, also called SLBP, which stands for stem-loop binding protein) binds specifically to a highly conserved hairpin structure located in the 3' UTR of the cell-cycle-dependent histone mRNAs. HBP consists of a minimal central RNA binding domain (RBD) flanked by an N- and C-terminal domain. The yeast three-hybrid system has been used to investigate the critical residues of the human HBP involved in the binding of its target hairpin structure. By means of negative selections followed by positive selections, we isolated mutant HBP species. Our results indicate tight relationships between the RBD and the N- and C-terminal domains.},
note = {0014-5793
Journal Article},
keywords = {Amino Acid Support, Amino Acid Sequence Amino Acid Substitution Base Sequence Carrier Proteins/chemistry/*genetics/*metabolism Histones/genetics Human Lac Operon/genetics Molecular Sequence Data Nucleic Acid Conformation Nucleotides/chemistry/genetics/metabolism RNA/chemistry/genetics/*metabolism RNA, ERIANI, Messenger/chemistry/genetics/metabolism RNA-Binding Proteins/chemistry/genetics/metabolism Recombinant Fusion Proteins/chemistry/genetics/metabolism Saccharomyces cerevisiae/*genetics Sequence Alignment Sequence Homology, Non-U.S. Gov't Two-Hybrid System Techniques beta-Galactosidase/genetics/metabolism, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Fender A, Geslain R, Eriani G, Giege R, Sissler M, Florentz C
A yeast arginine specific tRNA is a remnant aspartate acceptor Article de journal
Dans: Nucleic Acids Res, vol. 32, no. 17, p. 5076-5086, 2004, ISBN: 15452274, (1362-4962 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Arg/*chemistry/genetics/metabolism RNA, Asp/*chemistry/genetics/metabolism Saccharomyces cerevisiae/*genetics Sequence Alignment Support, ERIANI, ERIANI FLORENTZ GIEGE Aspartic Acid/metabolism Base Sequence *Evolution, FLORENTZ, Fungal/*chemistry/genetics/metabolism RNA, Molecular Molecular Sequence Data Point Mutation RNA, Non-U.S. Gov't, SISSLER, Transfer, Unité ARN
@article{,
title = {A yeast arginine specific tRNA is a remnant aspartate acceptor},
author = {A Fender and R Geslain and G Eriani and R Giege and M Sissler and C Florentz},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15452274},
isbn = {15452274},
year = {2004},
date = {2004-01-01},
journal = {Nucleic Acids Res},
volume = {32},
number = {17},
pages = {5076-5086},
abstract = {High specificity in aminoacylation of transfer RNAs (tRNAs) with the help of their cognate aminoacyl-tRNA synthetases (aaRSs) is a guarantee for accurate genetic translation. Structural and mechanistic peculiarities between the different tRNA/aaRS couples, suggest that aminoacylation systems are unrelated. However, occurrence of tRNA mischarging by non-cognate aaRSs reflects the relationship between such systems. In Saccharomyces cerevisiae, functional links between arginylation and aspartylation systems have been reported. In particular, it was found that an in vitro transcribed tRNAAsp is a very efficient substrate for ArgRS. In this study, the relationship of arginine and aspartate systems is further explored, based on the discovery of a fourth isoacceptor in the yeast genome, tRNA4Arg. This tRNA has a sequence strikingly similar to that of tRNAAsp but distinct from those of the other three arginine isoacceptors. After transplantation of the full set of aspartate identity elements into the four arginine isoacceptors, tRNA4Arg gains the highest aspartylation efficiency. Moreover, it is possible to convert tRNA4Arg into an aspartate acceptor, as efficient as tRNAAsp, by only two point mutations, C38 and G73, despite the absence of the major anticodon aspartate identity elements. Thus, cryptic aspartate identity elements are embedded within tRNA4Arg. The latent aspartate acceptor capacity in a contemporary tRNAArg leads to the proposal of an evolutionary link between tRNA4Arg and tRNAAsp genes.},
note = {1362-4962
Journal Article},
keywords = {Arg/*chemistry/genetics/metabolism RNA, Asp/*chemistry/genetics/metabolism Saccharomyces cerevisiae/*genetics Sequence Alignment Support, ERIANI, ERIANI FLORENTZ GIEGE Aspartic Acid/metabolism Base Sequence *Evolution, FLORENTZ, Fungal/*chemistry/genetics/metabolism RNA, Molecular Molecular Sequence Data Point Mutation RNA, Non-U.S. Gov't, SISSLER, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Ador L, Jaeger S, Geslain R, Martin F, Cavarelli J, Eriani G
Mutation and evolution of the magnesium-binding site of a class II aminoacyl-tRNA synthetase Article de journal
Dans: Biochemistry, vol. 43, no. 22, p. 7028-7037, 2004, ISBN: 15170340, (0006-2960 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Acylation Adenosine Triphosphate/metabolism Amino Acid Substitution Aspartate-tRNA Ligase/chemistry/genetics/*metabolism Aspartic Acid/metabolism Binding Sites Catalytic Domain Cell Death Combinatorial Chemistry Techniques Comparative Study *Evolution, Asp/metabolism Saccharomyces cerevisiae/*enzymology Support, ERIANI, Molecular Kinetics Magnesium/*metabolism Mutagenesis, Non-U.S. Gov't Transfection, Site-Directed Mutation/*genetics Peptide Library Protein Binding Protein Conformation RNA, Transfer, Unité ARN
@article{,
title = {Mutation and evolution of the magnesium-binding site of a class II aminoacyl-tRNA synthetase},
author = {L Ador and S Jaeger and R Geslain and F Martin and J Cavarelli and G Eriani},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15170340},
isbn = {15170340},
year = {2004},
date = {2004-01-01},
journal = {Biochemistry},
volume = {43},
number = {22},
pages = {7028-7037},
abstract = {Aminoacyl-tRNA synthetases contain one or three Mg(2+) ions in their catalytic sites. In addition to their role in ATP binding, these ions are presumed to play a role in catalysis by increasing the electropositivity of the alpha-phosphate and stabilizing the pentavalent transition state. In the class II aaRS, two highly conserved carboxylate residues have been shown to participate with Mg(2+) ions in binding and coordination. It is shown here that these carboxylate residues are absolutely required for the activity of Saccharomyces cerevisiae aspartyl-tRNA synthetase. Mutants of these residues exhibit pleiotropic effects on the kinetic parameters suggesting an effect at an early stage of the aminoacylation reaction, such as the binding of ATP, Mg(2+), aspartic acid, or the amino acid activation. Despite genetic selections in an APS-knockout yeast strain, we were unable to select a single active mutant of these carboxylate residues. Nevertheless, we isolated an intragenic suppressor from a combinatorial library. The active mutant showed a second substitution close to the first one, and exhibited a significant increase of the tRNA aminoacylation rate. Structural analysis suggests that the acceptor stem of the tRNA might be repositioned to give a more productive enzyme:tRNA complex. Thus, the initial defect of the activation reaction was compensated by a significant increase of the aminoacylation rate that led to cellular complementation.},
note = {0006-2960
Journal Article},
keywords = {Acylation Adenosine Triphosphate/metabolism Amino Acid Substitution Aspartate-tRNA Ligase/chemistry/genetics/*metabolism Aspartic Acid/metabolism Binding Sites Catalytic Domain Cell Death Combinatorial Chemistry Techniques Comparative Study *Evolution, Asp/metabolism Saccharomyces cerevisiae/*enzymology Support, ERIANI, Molecular Kinetics Magnesium/*metabolism Mutagenesis, Non-U.S. Gov't Transfection, Site-Directed Mutation/*genetics Peptide Library Protein Binding Protein Conformation RNA, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Sohm B, Frugier M, Brule H, Olszak K, Przykorska A, Florentz C
Towards understanding human mitochondrial leucine aminoacylation identity Article de journal
Dans: J Mol Biol, vol. 328, no. 5, p. 995-1010, 2003, ISBN: 12729737, (0022-2836 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Base Sequence Human In Vitro Leucine/*metabolism Leucine-tRNA Ligase/*metabolism Mitochondria/*metabolism Mitochondrial Diseases/genetics/metabolism Molecular Sequence Data Mutation Nucleic Acid Conformation RNA, ERIANI, FLORENTZ, FRUGIER, Leu/chemistry/genetics/*metabolism Recombinant Proteins/genetics/metabolism Solutions Substrate Specificity Support, Non-U.S. Gov't Variation (Genetics), Transfer, Unité ARN
@article{,
title = {Towards understanding human mitochondrial leucine aminoacylation identity},
author = {B Sohm and M Frugier and H Brule and K Olszak and A Przykorska and C Florentz},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12729737},
isbn = {12729737},
year = {2003},
date = {2003-01-01},
journal = {J Mol Biol},
volume = {328},
number = {5},
pages = {995-1010},
abstract = {Specific recognition of tRNAs by aminoacyl-tRNA synthetases is governed by sets of aminoacylation identity elements, well defined for numerous prokaryotic systems and eukaryotic cytosolic systems. Only restricted information is available for aminoacylation of human mitochondrial tRNAs, despite their particularities linked to the non-classical structures of the tRNAs and their involvement in a growing number of human neurodegenerative disorders linked to mutations in the corresponding tRNA genes. A major difficulty to be overcome is the preparation of active in vitro transcripts enabling a rational mutagenic analysis, as is currently performed for classical tRNAs. Here, structural and aminoacylation properties of in vitro transcribed tRNA(Leu(UUR)) are presented. Solution probing using a combination of enzymatic and chemical tools revealed only partial folding into an L-shaped structure, with an acceptor branch but with a floppy anticodon branch. Optimization of aminoacylation conditions allowed charging of up to 75% of molecules, showing that, despite its partially relaxed structure, in vitro transcribed tRNA(Leu(UUR)) is able to adapt to the synthetase. In addition, mutational analysis demonstrates that the discriminator base as well as residue A14 are important leucine identity elements. Thus, human mitochondrial leucylation is dependent on rules similar to those that apply in Escherichia coli. The impact of a subset of pathology-related mutations on aminoacylation and on tRNA structure, has been explored. These variants do not show significant structural rearrangements and either do not affect aminoacylation (mutations T3250C, T3271C, C3303T) or lead to marked effects. Interestingly, two variants with a mutation at the same position (A3243G and A3243T) lead to markedly different losses in aminoacylation efficiencies (tenfold and 300-fold, respectively).},
note = {0022-2836
Journal Article},
keywords = {Base Sequence Human In Vitro Leucine/*metabolism Leucine-tRNA Ligase/*metabolism Mitochondria/*metabolism Mitochondrial Diseases/genetics/metabolism Molecular Sequence Data Mutation Nucleic Acid Conformation RNA, ERIANI, FLORENTZ, FRUGIER, Leu/chemistry/genetics/*metabolism Recombinant Proteins/genetics/metabolism Solutions Substrate Specificity Support, Non-U.S. Gov't Variation (Genetics), Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Zhao M W, Hao R, Chen J F, Martin F, Eriani G, Wang E D
Enzymes assembled from Aquifex aeolicus and Escherichia coli leucyl-tRNA synthetases Article de journal
Dans: Biochemistry, vol. 42, no. 25, p. 7694-7700, 2003, ISBN: 12820878, (0006-2960 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Amino Acyl-tRNA Ligases/genetics/*metabolism Escherichia coli/*enzymology Evolution, ERIANI, Horizontal Heat Kinetics Mutation RNA, Leu/*metabolism Structure-Activity Relationship Support, Molecular Gene Transfer, Non-U.S. Gov't, Transfer, Unité ARN
@article{,
title = {Enzymes assembled from Aquifex aeolicus and Escherichia coli leucyl-tRNA synthetases},
author = {M W Zhao and R Hao and J F Chen and F Martin and G Eriani and E D Wang},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12820878},
isbn = {12820878},
year = {2003},
date = {2003-01-01},
journal = {Biochemistry},
volume = {42},
number = {25},
pages = {7694-7700},
abstract = {Aquifex aeolicus alphabeta-LeuRS is the only known heterodimeric LeuRS, while Escherichia coli LeuRS is a canonical monomeric enzyme. By using the genes encoding A. aeolicus and E. coli LeuRS as PCR templates, the genes encoding the alpha and beta subunits from A. aeolicus alphabeta-LeuRS and the equivalent amino- and carboxy-terminal parts of E. coli LeuRS (identified as alpha' and beta') were amplified and recombined using suitable plasmids. These recombinant plasmids were transformed or cotransformed into E. coli to produce five monomeric and five heterodimeric LeuRS mutants. Seven of these were successfully overexpressed in vivo and purified, while three dimeric mutants with the beta' part of E. coli LeuRS were not successfully expressed. The seven purified mutants catalyzed amino acid activation, although several exhibited reduced aminoacylation properties. Removal of the last 36 residues of the alpha subunit of the A. aeolicus enzyme was determined to be deleterious for tRNA charging. Indeed, subunit exchange showed that the cross-species-specific recognition of A. aeolicus tRNA(Leu) occurs at the alpha subunit. None of the mixed E. coli-A. aeolicus enzymes were as thermostable as the native alphabeta-LeuRS. However, the fusion of the two alpha and beta peptides from A. aeolicus as a single chain analogous to canonical LeuRS resulted in a product more resistant to heat denaturation than the original enzyme.},
note = {0006-2960
Journal Article},
keywords = {Amino Acyl-tRNA Ligases/genetics/*metabolism Escherichia coli/*enzymology Evolution, ERIANI, Horizontal Heat Kinetics Mutation RNA, Leu/*metabolism Structure-Activity Relationship Support, Molecular Gene Transfer, Non-U.S. Gov't, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Geslain R, Martin F, Camasses A, Eriani G
A yeast knockout strain to discriminate between active and inactive tRNA molecules Article de journal
Dans: Nucleic Acids Res, vol. 31, no. 16, p. 4729-4737, 2003, ISBN: 12907713, (1362-4962 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Amino Acyl-tRNA Ligases/metabolism Arginine/genetics/metabolism Base Sequence Blotting, Arg/chemistry/*genetics/metabolism Saccharomyces cerevisiae/*genetics Support, ERIANI, Molecular Hydrogen-Ion Concentration Molecular Sequence Data Mutagenesis, Non-U.S. Gov't, Northern Cloning, Site-Directed Mutation Nucleic Acid Conformation RNA, Transfer, Unité ARN
@article{,
title = {A yeast knockout strain to discriminate between active and inactive tRNA molecules},
author = {R Geslain and F Martin and A Camasses and G Eriani},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12907713},
isbn = {12907713},
year = {2003},
date = {2003-01-01},
journal = {Nucleic Acids Res},
volume = {31},
number = {16},
pages = {4729-4737},
abstract = {Here we report the construction of a yeast genetic screen designed to identify essential residues in tRNA(Arg). The system consists of a tRNA(Arg) knockout strain and a set of vectors designed to rescue and select for variants of tRNA(Arg). By plasmid shuffling we selected inactive tRNA mutants that were further analyzed by northern blotting. The mutational analysis focused on the tRNA D and anticodon loops that contact the aminoacyl-tRNA synthetase. The anticodon triplet was excluded from the analysis because of its role in decoding the Arg codons. Most of the inactivating mutations are residues involved in tertiary interactions. These mutations had dramatic effects on tRNA(Arg) abundance. Other inactivating mutations were located in the anticodon loop, where they did not affect transcription and aminoacylation but probably altered interaction with the translation machinery. No lethal effects were observed when residues 16, 20 and 38 were individually mutated, despite the fact that they are involved in sequence-specific interactions with the aminoacyl-tRNA synthetase. However, the steady-state levels of the aminoacylated forms of U20A and U20G were decreased by a factor of 3.5-fold in vivo. This suggests that, unlike in the Escherichia coli tRNA(Arg):ArgRS system where residue 20 (A) is a major identity element, in yeast this position is of limited consequence.},
note = {1362-4962
Journal Article},
keywords = {Amino Acyl-tRNA Ligases/metabolism Arginine/genetics/metabolism Base Sequence Blotting, Arg/chemistry/*genetics/metabolism Saccharomyces cerevisiae/*genetics Support, ERIANI, Molecular Hydrogen-Ion Concentration Molecular Sequence Data Mutagenesis, Non-U.S. Gov't, Northern Cloning, Site-Directed Mutation Nucleic Acid Conformation RNA, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Geslain R, Bey G, Cavarelli J, Eriani G
Limited set of amino acid residues in a class Ia aminoacyl-tRNA synthetase is crucial for tRNA binding Article de journal
Dans: Biochemistry, vol. 42, no. 51, p. 15092-15101, 2003, ISBN: 14690419, (0006-2960 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Acylation Alanine/genetics Amino Acids/*chemistry/genetics Anticodon/chemistry/genetics Arginine-tRNA Ligase/*chemistry/classification/genetics Binding Sites/genetics Genes, Arg/*chemistry Saccharomyces cerevisiae/enzymology/genetics/growth & development Saccharomyces cerevisiae Proteins/*chemistry/*genetics Support, ERIANI, Lethal Mutagenesis, Non-U.S. Gov't, Site-Directed Protein Binding/genetics Protein Structure, Tertiary/genetics RNA, Transfer, Unité ARN
@article{,
title = {Limited set of amino acid residues in a class Ia aminoacyl-tRNA synthetase is crucial for tRNA binding},
author = {R Geslain and G Bey and J Cavarelli and G Eriani},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14690419},
isbn = {14690419},
year = {2003},
date = {2003-01-01},
journal = {Biochemistry},
volume = {42},
number = {51},
pages = {15092-15101},
abstract = {The aim of this work was to characterize crucial amino acids for the aminoacylation of tRNA(Arg) by yeast arginyl-tRNA synthetase. Alanine mutagenesis was used to probe all the side chain mediated interactions that occur between tRNA(Arg2)(ICG) and ArgRS. The effects of the substitutions were analyzed in vivo in an ArgRS-knockout strain and in vitro by measuring the aminoacylation efficiencies for two distinct tRNA(Arg) isoacceptors. Nine mutants that generate lethal phenotypes were identified, suggesting that only a limited set of side chain mediated interactions is essential for tRNA recognition. The majority of the lethal mutants was mapped to the anticodon binding domain of ArgRS, a helix bundle that is characteristic for class Ia synthetases. The alanine mutations induce drastic decreases in the tRNA charging rates, which is correlated with a loss in affinity in the catalytic site for ATP. One of those lethal mutations corresponds to an Arg residue that is strictly conserved in all class Ia synthetases. In the known crystallographic structures of complexes of tRNAs and class Ia synthetases, this invariant Arg residue stabilizes the idiosyncratic conformation of the anticodon loop. This paper also highlights the crucial role of the tRNA and enzyme plasticity upon binding. Divalent ions are also shown to contribute to the induced fit process as they may stabilize the local tRNA-enzyme interface. Furthermore, one lethal phenotype can be reverted in the presence of high Mg(2+) concentrations. In contrast with the bacterial system, in yeast arginyl-tRNA synthetase, no lethal mutation has been found in the ArgRS specific domain recognizing the Dhu-loop of the tRNA(Arg). Mutations in this domain have no effects on tRNA(Arg) aminoacylation, thus confirming that Saccharomyces cerevisiae and other fungi belong to a distinct class of ArgRS.},
note = {0006-2960
Journal Article},
keywords = {Acylation Alanine/genetics Amino Acids/*chemistry/genetics Anticodon/chemistry/genetics Arginine-tRNA Ligase/*chemistry/classification/genetics Binding Sites/genetics Genes, Arg/*chemistry Saccharomyces cerevisiae/enzymology/genetics/growth & development Saccharomyces cerevisiae Proteins/*chemistry/*genetics Support, ERIANI, Lethal Mutagenesis, Non-U.S. Gov't, Site-Directed Protein Binding/genetics Protein Structure, Tertiary/genetics RNA, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Xu M G, Chen J F, Martin F, Zhao M W, Eriani G, Wang E D
Leucyl-tRNA synthetase consisting of two subunits from hyperthermophilic bacteria Aquifex aeolicus Article de journal
Dans: J Biol Chem, vol. 277, no. 44, p. 41590-41596, 2002, ISBN: 12196521, (0021-9258 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Bacteria/*enzymology Binding Sites Circular Dichroism Cloning, ERIANI, Leu Support, Molecular Enzyme Stability Escherichia coli/enzymology Kinetics Leucine-tRNA Ligase/*chemistry/genetics/isolation & purification Protein Subunits RNA, Non-U.S. Gov't, Transfer, Unité ARN
@article{,
title = {Leucyl-tRNA synthetase consisting of two subunits from hyperthermophilic bacteria Aquifex aeolicus},
author = {M G Xu and J F Chen and F Martin and M W Zhao and G Eriani and E D Wang},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12196521},
isbn = {12196521},
year = {2002},
date = {2002-01-01},
journal = {J Biol Chem},
volume = {277},
number = {44},
pages = {41590-41596},
abstract = {In a hyperthermophilic bacterium, Aquifex aeolicus, leucyl-tRNA synthetase (LeuRS) consists of two non-identical polypeptide subunits (alpha and beta), different from the canonical LeuRS, which has a single polypeptide chain. By PCR, using genome DNA of A. aeolicus as a template, genes encoding the alpha and beta subunits were amplified and cloned in Escherichia coli. The alpha subunit could not be expressed stably in vivo, whereas the beta subunit was overproduced and purified by a simple procedure. The beta subunit was inactive in catalysis but was able to bind tRNA(Leu). Interestingly, the heterodimer alphabeta-LeuRS could be overproduced in E. coli cells containing both genes and was purified to 95% homogeneity as a hybrid dimer. The kinetics of A. aeolicus LeuRS in pre-steady and steady states and cross-recognition of LeuRS and tRNA(Leu) from A. aeolicus and E. coli were studied. Magnesium concentration, pH value, and temperature aminoacylation optima were determined to be 12 mm, 7.8, and 70 degrees C, respectively. Under optimal conditions, A. aeolicus alphabeta-LeuRS is stable up to 65 degrees C.},
note = {0021-9258
Journal Article},
keywords = {Bacteria/*enzymology Binding Sites Circular Dichroism Cloning, ERIANI, Leu Support, Molecular Enzyme Stability Escherichia coli/enzymology Kinetics Leucine-tRNA Ligase/*chemistry/genetics/isolation & purification Protein Subunits RNA, Non-U.S. Gov't, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Lescure A, Allmang C, Yamada K, Carbon P, Krol A
cDNA cloning, expression pattern and RNA binding analysis of human selenocysteine insertion sequence (SECIS) binding protein 2 Article de journal
Dans: Gene, vol. 291, no. 1-2, p. 279-285, 2002, ISBN: 12095701, (0378-1119 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Amino Acid Support, Amino Acid Sequence Blotting, Complementary/chemistry/genetics Female Gene Expression Human Male Molecular Sequence Data Peptide Elongation Factors/metabolism Protein Binding RNA/*metabolism RNA, DNA Sequence Homology, ERIANI, LESCURE, Messenger/genetics/metabolism RNA-Binding Proteins/genetics/*metabolism Sequence Alignment Sequence Analysis, Molecular DNA, Non-U.S. Gov't, Northern Cloning, Unité ARN
@article{,
title = {cDNA cloning, expression pattern and RNA binding analysis of human selenocysteine insertion sequence (SECIS) binding protein 2},
author = {A Lescure and C Allmang and K Yamada and P Carbon and A Krol},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12095701},
isbn = {12095701},
year = {2002},
date = {2002-01-01},
journal = {Gene},
volume = {291},
number = {1-2},
pages = {279-285},
abstract = {Selenocysteine and selenoprotein synthesis require a complex molecular machinery in mammals. Among the key players is the RNA-protein complex formed by the selenocysteine insertion sequence (SECIS) binding protein (SBP2) and the SECIS element, an RNA hairpin in the 3' untranslated regions of selenoprotein messenger RNAs (mRNAs). We have isolated the DNA complementary to mRNA of the human SBP2, enabling us to establish that it differs from a previously reported human SBP2-like protein. Examination of the expression pattern revealed that the human SBP2 protein is encoded by a 4 kb long mRNA that is over-expressed in testis. Compared to the rat SBP2 sequence, the human SBP2 protein displays two highly conserved domains with 92 and 95% amino acid identity, the latter one containing the RNA binding domain. The inter-domain section carries 55% sequence identity, the remainder of the SBP2 sequences showing about 65% identity, values lower than expected for two mammalian proteins. Interestingly, we could show that the binding of human SBP2 to the SECIS RNA is stimulated by the selenoprotein-specialized elongation translation factor mSelB/eEFsec.},
note = {0378-1119
Journal Article},
keywords = {Amino Acid Support, Amino Acid Sequence Blotting, Complementary/chemistry/genetics Female Gene Expression Human Male Molecular Sequence Data Peptide Elongation Factors/metabolism Protein Binding RNA/*metabolism RNA, DNA Sequence Homology, ERIANI, LESCURE, Messenger/genetics/metabolism RNA-Binding Proteins/genetics/*metabolism Sequence Alignment Sequence Analysis, Molecular DNA, Non-U.S. Gov't, Northern Cloning, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Allmang C, Carbon P, Krol A
The SBP2 and 15.5 kD/Snu13p proteins share the same RNA binding domain: identification of SBP2 amino acids important to SECIS RNA binding Article de journal
Dans: RNA, vol. 8, no. 10, p. 1308-1318, 2002, ISBN: 12403468, (1355-8382 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Amino Acid Spliceosomes/metabolism Structural Homology, Amino Acid Sequence Amino Acid Substitution Binding Sites Human Molecular Sequence Data Mutation Nucleic Acid Conformation RNA, ERIANI, Non-U.S. Gov't, Protein Support, Small Nuclear/chemistry/*metabolism RNA-Binding Proteins/genetics/*metabolism Ribonucleoproteins, Small Nuclear/genetics/*metabolism Selenocysteine/metabolism Sequence Homology, Unité ARN
@article{,
title = {The SBP2 and 15.5 kD/Snu13p proteins share the same RNA binding domain: identification of SBP2 amino acids important to SECIS RNA binding},
author = {C Allmang and P Carbon and A Krol},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12403468},
isbn = {12403468},
year = {2002},
date = {2002-01-01},
journal = {RNA},
volume = {8},
number = {10},
pages = {1308-1318},
abstract = {Selenoprotein synthesis in eukaryotes requires the selenocysteine insertion sequence (SECIS) RNA, a hairpin in the 3' untranslated region of selenoprotein mRNAs. The SECIS RNA is recognized by the SECIS-binding protein 2 (SBP2), which is a key player in this specialized translation machinery. The objective of this work was to obtain structural insight into the SBP2-SECIS RNA complex. Multiple sequence alignment revealed that SBP2 and the U4 snRNA-binding protein 15.5 kD/Snu13p share the same RNA binding domain of the L7A/L30 family, also found in the box H/ACA snoRNP protein Nhp2p and several ribosomal proteins. In corollary, we have detected a similar secondary structure motif in the SECIS and U4 RNAs. Combining the data of the crystal structure of the 15.5 kD-U4 snRNA complex, and the SBP2/15.5 kD sequence similarities, we designed a structure-guided strategy predicting 12 SBP2 amino acids that should be critical for SECIS RNA binding. Alanine substitution of these amino acids followed by gel shift assays of the SBP2 mutant proteins identified four residues whose mutation severely diminished or abolished SECIS RNA binding, the other eight provoking intermediate down effects. In addition to identifying key amino acids for SECIS recognition by SBP2, our findings led to the proposal that some of the recognition principles governing the 15.5 kD-U4 snRNA interaction must be similar in the SBP2-SECIS RNA complex.},
note = {1355-8382
Journal Article},
keywords = {Amino Acid Spliceosomes/metabolism Structural Homology, Amino Acid Sequence Amino Acid Substitution Binding Sites Human Molecular Sequence Data Mutation Nucleic Acid Conformation RNA, ERIANI, Non-U.S. Gov't, Protein Support, Small Nuclear/chemistry/*metabolism RNA-Binding Proteins/genetics/*metabolism Ribonucleoproteins, Small Nuclear/genetics/*metabolism Selenocysteine/metabolism Sequence Homology, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
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}
}
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}
}
Wu J F, Wang E D, Wang Y L, Eriani G, Gangloff J
Gene cloning, overproduction and purification of Escherichia coli tRNA(Arg2). Article de journal
Dans: Acta Bioch Bioph Sin, vol. 31, no. 3, p. 226-232, 1999, ISBN: 12136168.
Résumé | Liens | BibTeX | Étiquettes: ERIANI, Unité ARN
@article{,
title = {Gene cloning, overproduction and purification of Escherichia coli tRNA(Arg2).},
author = {J F Wu and E D Wang and Y L Wang and G Eriani and J Gangloff},
url = {http://www.ncbi.nlm.nih.gov/pubmed/12136168},
isbn = {12136168},
year = {1999},
date = {1999-01-01},
journal = {Acta Bioch Bioph Sin},
volume = {31},
number = {3},
pages = {226-232},
abstract = {A synthetic gene encoding Escherichia coli tRNA(Arg)(2) was inserted in a plasmid under the control of an isopropyl-beta, D-thiogalactopyranoside (IPTG)-inducible promotor, pTrc99B. In E.coli MT102 transformed by the above plasmid containing the target gene. TRNA(Arg)(2) was overproduced up to 30 fold of that of the host. In the transformant the quantity contained tRNA(Arg) increased 10 times and was 70% of the total tRNA. The tRNA(Arg)(2) was purified to 88% homogeneity by passing through a DEAE-Sephacel column, and then was purified by benzyl-DEAE cellulose column chromatography to a purity of 99% with an arginylation activity of 1 600 pmole/A(260) unit. Eighteen milligrams of tRNA(Arg)(2) could be obtained from 40 mg total tRNA which was obtained from four liters of overnight culture, and the yield of the purification was 62%. The accurate kinetic constants of aminoacylation of tRNA(Arg)(2) catalyzed by arginyl-tRNA synthetase were comparable with that of tRNA(Arg) from Sigma.},
keywords = {ERIANI, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
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}
}
Martin F, Sharples G J, Lloyd R G, Eiler S, Moras D, Gangloff J, Eriani G
Characterization of a thermosensitive Escherichia coli aspartyl-tRNA synthetase mutant Article de journal
Dans: J Bacteriol, vol. 179, no. 11, p. 3691-3696, 1997, ISBN: 9171418, (0021-9193 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Aspartate-tRNA Ligase/*genetics Escherichia coli/*genetics Models, ERIANI, Molecular Mutation Structure-Activity Relationship Support, Non-U.S. Gov't Temperature, Unité ARN
@article{,
title = {Characterization of a thermosensitive Escherichia coli aspartyl-tRNA synthetase mutant},
author = {F Martin and G J Sharples and R G Lloyd and S Eiler and D Moras and J Gangloff and G Eriani},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9171418},
isbn = {9171418},
year = {1997},
date = {1997-01-01},
journal = {J Bacteriol},
volume = {179},
number = {11},
pages = {3691-3696},
abstract = {The Escherichia coli tls-1 strain carrying a mutated aspS gene (coding for aspartyl-tRNA synthetase), which causes a temperature-sensitive growth phenotype, was cloned by PCR, sequenced, and shown to contain a single mutation resulting in substitution by serine of the highly conserved proline 555, which is located in motif 3. When an aspS fragment spanning the codon for proline 555 was transformed into the tls-1 strain, it was shown to restore the wild-type phenotype via homologous recombination with the chromosomal tls-1 allele. The mutated AspRS purified from an overproducing strain displayed marked temperature sensitivity, with half-life values of 22 and 68 min (at 42 degrees C), respectively, for tRNA aminoacylation and ATP/PPi exchange activities. Km values for aspartic acid, ATP, and tRNA(Asp) did not significantly differ from those of the native enzyme; thus, mutation Pro555Ser lowers the stability of the functional configuration of both the acylation and the amino acid activation sites but has no significant effect on substrate binding. This decrease in stability appears to be related to a conformational change, as shown by gel filtration analysis. Structural data strongly suggest that the Pro555Ser mutation lowers the stability of the Lys556 and Thr557 positions, since these two residues, as shown by the crystallographic structure of the enzyme, are involved in the active site and in contacts with the tRNA acceptor arm, respectively.},
note = {0021-9193
Journal Article},
keywords = {Aspartate-tRNA Ligase/*genetics Escherichia coli/*genetics Models, ERIANI, Molecular Mutation Structure-Activity Relationship Support, Non-U.S. Gov't Temperature, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Martin F, Reinbolt J, Dirheimer G, Gangloff J, Eriani G
Selection of tRNA(Asp) amber suppressor mutants having alanine, arginine, glutamine, and lysine identity Article de journal
Dans: RNA, vol. 2, no. 9, p. 919-927, 1996, ISBN: 8809018, (1355-8382 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Alanine/genetics Arginine/genetics Base Sequence Escherichia coli/genetics Genes, Asp/*genetics *Selection (Genetics) Support, ERIANI, Genetic, Genetic Molecular Sequence Data *Mutation RNA, Non-U.S. Gov't *Suppression, Suppressor Glutamine/genetics Lysine/genetics Models, Transfer, Unité ARN
@article{,
title = {Selection of tRNA(Asp) amber suppressor mutants having alanine, arginine, glutamine, and lysine identity},
author = {F Martin and J Reinbolt and G Dirheimer and J Gangloff and G Eriani},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8809018},
isbn = {8809018},
year = {1996},
date = {1996-01-01},
journal = {RNA},
volume = {2},
number = {9},
pages = {919-927},
abstract = {Elements that confer identity to a tRNA in the cellular environment, where all aminoacyl-tRNA synthetases are competing for substrates, may be delineated by in vivo experiments using suppressor tRNAs. Here we describe the selection of active Escherichia coli tRNAAsp amber mutants and analyze their identity. Starting from a library containing randomly mutated tRNA(CUA)Asp genes, we isolated four amber suppressors presenting either lysine, alanine, or glutamine activity. Two of them, presenting mainly alanine or lysine activity, were further submitted to a second round of mutagenesis selection in order to improve their efficiency of suppression. Eleven suppressors were isolated, each containing two or three mutations. Ten presented identities of the two parental mutants, whereas one had switched from lysine to arginine identity. Analysis of the different mutants revealed (or confirmed for some nucleotides) their role as positive and/or negative determinants in AlaRS, LysRS, and ArgRS recognition. More generally, it appears that tRNAAsp presents identity characteristics closely related to those of tRNALys, as well as a structural basis for acquiring alanine or arginine identity upon moderate mutational changes; these consist of addition or suppression of the corresponding positive or negative determinants, as well as tertiary interactions. Failure to isolate aspartic acid-inserting suppressors is probably due to elimination of the important G34 identity element and its replacement by an antideterminant when changing the anticodon of the tRNAAsp to the CUA triplet.},
note = {1355-8382
Journal Article},
keywords = {Alanine/genetics Arginine/genetics Base Sequence Escherichia coli/genetics Genes, Asp/*genetics *Selection (Genetics) Support, ERIANI, Genetic, Genetic Molecular Sequence Data *Mutation RNA, Non-U.S. Gov't *Suppression, Suppressor Glutamine/genetics Lysine/genetics Models, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Martin F, Reinbolt J, Dirheimer G, Gangloff J, Eriani G
Selection of tRNA(Asp) amber suppressor mutants having alanine, arginine, glutamine, and lysine identity Article de journal
Dans: RNA, vol. 2, no. 9, p. 919-927, 1996, ISBN: 8809018, (1355-8382 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Alanine/genetics Arginine/genetics Base Sequence Escherichia coli/genetics Genes, Asp/*genetics *Selection (Genetics) Support, ERIANI, Genetic, Genetic Molecular Sequence Data *Mutation RNA, Non-U.S. Gov't *Suppression, Suppressor Glutamine/genetics Lysine/genetics Models, Transfer, Unité ARN
@article{,
title = {Selection of tRNA(Asp) amber suppressor mutants having alanine, arginine, glutamine, and lysine identity},
author = {F Martin and J Reinbolt and G Dirheimer and J Gangloff and G Eriani},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8809018},
isbn = {8809018},
year = {1996},
date = {1996-01-01},
journal = {RNA},
volume = {2},
number = {9},
pages = {919-927},
abstract = {Elements that confer identity to a tRNA in the cellular environment, where all aminoacyl-tRNA synthetases are competing for substrates, may be delineated by in vivo experiments using suppressor tRNAs. Here we describe the selection of active Escherichia coli tRNAAsp amber mutants and analyze their identity. Starting from a library containing randomly mutated tRNA(CUA)Asp genes, we isolated four amber suppressors presenting either lysine, alanine, or glutamine activity. Two of them, presenting mainly alanine or lysine activity, were further submitted to a second round of mutagenesis selection in order to improve their efficiency of suppression. Eleven suppressors were isolated, each containing two or three mutations. Ten presented identities of the two parental mutants, whereas one had switched from lysine to arginine identity. Analysis of the different mutants revealed (or confirmed for some nucleotides) their role as positive and/or negative determinants in AlaRS, LysRS, and ArgRS recognition. More generally, it appears that tRNAAsp presents identity characteristics closely related to those of tRNALys, as well as a structural basis for acquiring alanine or arginine identity upon moderate mutational changes; these consist of addition or suppression of the corresponding positive or negative determinants, as well as tertiary interactions. Failure to isolate aspartic acid-inserting suppressors is probably due to elimination of the important G34 identity element and its replacement by an antideterminant when changing the anticodon of the tRNAAsp to the CUA triplet.},
note = {1355-8382
Journal Article},
keywords = {Alanine/genetics Arginine/genetics Base Sequence Escherichia coli/genetics Genes, Asp/*genetics *Selection (Genetics) Support, ERIANI, Genetic, Genetic Molecular Sequence Data *Mutation RNA, Non-U.S. Gov't *Suppression, Suppressor Glutamine/genetics Lysine/genetics Models, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Giege R, Florentz C, Kern D, Gangloff J, Eriani G, Moras D
Aspartate identity of transfer RNAs Article de journal
Dans: Biochimie, vol. 78, no. 7, p. 605-623, 1996, ISBN: 8955904, (0300-9084 Journal Article Review Review, Tutorial).
Résumé | Liens | BibTeX | Étiquettes: Asp/*chemistry Saccharomyces cerevisiae Structure-Activity Relationship Support, Aspartate-tRNA Ligase/chemistry/metabolism Aspartic Acid/analysis Base Sequence Escherichia coli Models, ERIANI, FLORENTZ, Molecular Molecular Sequence Data Nucleic Acid Conformation RNA, Non-U.S. Gov't Thermus thermophilus, Transfer, Unité ARN
@article{,
title = {Aspartate identity of transfer RNAs},
author = {R Giege and C Florentz and D Kern and J Gangloff and G Eriani and D Moras},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8955904},
isbn = {8955904},
year = {1996},
date = {1996-01-01},
journal = {Biochimie},
volume = {78},
number = {7},
pages = {605-623},
abstract = {Structure/function relationships accounting for specific tRNA charging by class II aspartyl-tRNA synthetases from Saccharomyces cerevisiae, Escherichia coli and Thermus thermophilus are reviewed. Effects directly linked to tRNA features are emphasized and aspects about synthetase contribution in expression of tRNA(Asp) identity are also covered. Major identity nucleotides conferring aspartate specificity to yeast, E coli and T thermophilus tRNAs comprise G34, U35, C36, C38 and G73, a set of nucleotides conserved in tRNA(Asp) molecules of other biological origin. Aspartate specificity can be enhanced by negative discrimination preventing, eg mischarging of native yeast tRNA(Asp by yeast arginyl-tRNA synthetase. In the yeast system crystallography shows that identity nucleotides are in contact with identity amino acids located in the catalytic and anticodon binding domains of the synthetase. Specificity of RNA/protein interaction involves a conformational change of the tRNA that optimizes the H-bonding potential of the identity signals on both partners of the complex. Mutation of identity nucleotides leads to decreased aspartylation efficiencies accompanied by a loss of specific H-bonds and an altered adaptation of tRNA on the synthetase. Species-specific characteristics of aspartate systems are the number, location and nature of minor identity signals. These features and the structural variations in aspartate tRNAs and synthetases are correlated with mechanistic differences in the aminoacylation reactions catalyzed by the various aspartyl-tRNA synthetases. The reality of the aspartate identity set is verified by its functional expression in a variety of RNA frameworks. Inversely a number of identities can be expressed within a tRNA(Asp) framework. From this emerged the concept of the RNA structural frameworks underlying expression of identities which is illustrated with data obtained with engineered tRNAs. Efficient aspartylation of minihelices is explained by the primordial role of G73. From this and other considerations it is suggested that aspartate identity appeared early in the history of tRNA aminoacylation systems.},
note = {0300-9084
Journal Article
Review
Review, Tutorial},
keywords = {Asp/*chemistry Saccharomyces cerevisiae Structure-Activity Relationship Support, Aspartate-tRNA Ligase/chemistry/metabolism Aspartic Acid/analysis Base Sequence Escherichia coli Models, ERIANI, FLORENTZ, Molecular Molecular Sequence Data Nucleic Acid Conformation RNA, Non-U.S. Gov't Thermus thermophilus, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Boeglin M, Dock-Brégeon A C, Eriani G, Gangloff J, Ruff M, Poterszman A, Thierry J C, Moras D
Crystallization of Escherichia coli aspartyl-tRNA synthetase in its free state and in a complex with yeast tRNA(Asp). Article de journal
Dans: Acta Crystallogr D Biol Crystallogr, vol. 52, no. Pt 1, p. 211-214, 1996, ISBN: 15299749.
Résumé | Liens | BibTeX | Étiquettes: ERIANI, Unité ARN
@article{,
title = {Crystallization of Escherichia coli aspartyl-tRNA synthetase in its free state and in a complex with yeast tRNA(Asp).},
author = {M Boeglin and A C Dock-Brégeon and G Eriani and J Gangloff and M Ruff and A Poterszman and J C Thierry and D Moras},
url = {http://www.ncbi.nlm.nih.gov/pubmed/15299749},
isbn = {15299749},
year = {1996},
date = {1996-01-01},
journal = {Acta Crystallogr D Biol Crystallogr},
volume = {52},
number = {Pt 1},
pages = {211-214},
abstract = {Overexpressed dimeric E. coli aspartyl-tRNA synthetase (AspRS) has been crystallized in its free state and complexed with yeast tRNA(Asp). Triclinic crystals of the enzyme alone (a = 104.4},
keywords = {ERIANI, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Wang E D, Huang Y W, Wang Y L, Eriani G, Gangloff J
The role of Lys378 and Lys381 on the enzyme activity of E. coli arginyl-tRNA synthetase. Article de journal
Dans: Acta Bioch Bioph Sin, vol. 27, p. 37-43, 1995.
Liens | BibTeX | Étiquettes: ERIANI, Unité ARN
@article{,
title = {The role of Lys378 and Lys381 on the enzyme activity of E. coli arginyl-tRNA synthetase.},
author = {E D Wang and Y W Huang and Y L Wang and G Eriani and J Gangloff},
url = {none},
year = {1995},
date = {1995-01-01},
journal = {Acta Bioch Bioph Sin},
volume = {27},
pages = {37-43},
keywords = {ERIANI, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Martin F, Eriani G, Reinbolt J, Dirheimer G, Gangloff J
Genetic selection for active E.coli amber tRNA(Asn) exclusively led to glutamine inserting suppressors Article de journal
Dans: Nucleic Acids Res, vol. 23, no. 5, p. 779-784, 1995, ISBN: 7708493, (0305-1048 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Asn/*genetics Support, Base Sequence Escherichia coli/*genetics *Genes, ERIANI, Insertional RNA, Non-U.S. Gov't, Suppressor Glutamine/*genetics Molecular Sequence Data Mutagenesis, Transfer, Unité ARN
@article{,
title = {Genetic selection for active E.coli amber tRNA(Asn) exclusively led to glutamine inserting suppressors},
author = {F Martin and G Eriani and J Reinbolt and G Dirheimer and J Gangloff},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=7708493},
isbn = {7708493},
year = {1995},
date = {1995-01-01},
journal = {Nucleic Acids Res},
volume = {23},
number = {5},
pages = {779-784},
abstract = {Suppressor tRNAs are useful tools for determining identity elements which define recognition of tRNAs in vivo by their cognate aminoacyl-tRNA synthetases. This study was aimed at the isolation of active amber tRNA(Asn). Nineteen mutated tRNA(Asn)CUA having amber suppressor activity were selected by an in vivo genetic screen, and all exclusively inserted glutamine. From analysis of the different mutations it is concluded that glutamine accepting activity was obtained upon reducing the interaction strength between the first base pair of the tRNA(Asn)CUA by direct or indirect effects. Failure to isolate tRNA(Asn)CUA suppressors charged with asparagine as well as other evolutionary related amino acids is discussed.},
note = {0305-1048
Journal Article},
keywords = {Asn/*genetics Support, Base Sequence Escherichia coli/*genetics *Genes, ERIANI, Insertional RNA, Non-U.S. Gov't, Suppressor Glutamine/*genetics Molecular Sequence Data Mutagenesis, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Eriani G, Cavarelli J, Martin F, Ador L, Rees B, Thierry J C, Gangloff J, Moras D
The class II aminoacyl-tRNA synthetases and their active site: evolutionary conservation of an ATP binding site Article de journal
Dans: J Mol Evol, vol. 40, no. 5, p. 499-508, 1995, ISBN: 7783225, (0022-2844 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Amino Acid Species Specificity Structure-Activity Relationship, Asp/metabolism Saccharomyces cerevisiae/enzymology Sequence Alignment Sequence Homology, ERIANI, Molecular Molecular Sequence Data Protein Binding Protein Conformation RNA, Transfer, Transfer/metabolism RNA, Unité ARN
@article{,
title = {The class II aminoacyl-tRNA synthetases and their active site: evolutionary conservation of an ATP binding site},
author = {G Eriani and J Cavarelli and F Martin and L Ador and B Rees and J C Thierry and J Gangloff and D Moras},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=7783225},
isbn = {7783225},
year = {1995},
date = {1995-01-01},
journal = {J Mol Evol},
volume = {40},
number = {5},
pages = {499-508},
abstract = {Previous sequence analyses have suggested the existence of two distinct classes of aminoacyl-tRNA synthetase. The partition was established on the basis of exclusive sets of sequence motifs (Eriani et al. [1990] Nature 347:203-306). X-ray studies have now well defined the structural basis of the two classes: the class I enzymes share with dehydrogenases and kinases the classic nucleotide binding fold called the Rossmann fold, whereas the class II enzymes possess a different fold, not found elsewhere, built around a six-stranded antiparallel beta-sheet. The two classes of synthetases catalyze the same global reaction that is the attachment of an amino acid to the tRNA, but differ as to where on the terminal adenosine of the tRNA the amino acid is placed: class I enzymes act on the 2' hydroxyl whereas the class II enzymes prefer the 3' hydroxyl group. The three-dimensional structure of aspartyl-tRNA synthetase from yeast, a typical class II enzyme, is described here, in relation to its function. The crucial role of the sequence motifs in substrate binding and enzyme structure is high-lighted. Overall these results underline the existence of an intimate evolutionary link between the aminoacyl-tRNA synthetases, despite their actual structural diversity.},
note = {0022-2844
Journal Article},
keywords = {Amino Acid Species Specificity Structure-Activity Relationship, Asp/metabolism Saccharomyces cerevisiae/enzymology Sequence Alignment Sequence Homology, ERIANI, Molecular Molecular Sequence Data Protein Binding Protein Conformation RNA, Transfer, Transfer/metabolism RNA, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Wang E D, Gu W L, Wang Y L, Eriani G, Gangloff J
Lys306 of E. coli arginyl-tRNA synthetase is necessary for the activity of this enzyme. Article de journal
Dans: Acta Bioch Bioph Sin, vol. 27, p. 123-128, 1995.
Liens | BibTeX | Étiquettes: ERIANI, Unité ARN
@article{,
title = {Lys306 of E. coli arginyl-tRNA synthetase is necessary for the activity of this enzyme.},
author = {E D Wang and W L Gu and Y L Wang and G Eriani and J Gangloff},
url = {none},
year = {1995},
date = {1995-01-01},
journal = {Acta Bioch Bioph Sin},
volume = {27},
pages = {123-128},
keywords = {ERIANI, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Frugier M, Soll D, Giege R, Florentz C
Identity switches between tRNAs aminoacylated by class I glutaminyl- and class II aspartyl-tRNA synthetases Article de journal
Dans: Biochemistry, vol. 33, no. 33, p. 9912-9921, 1994, ISBN: 8060999, (0006-2960 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Acylation Aspartate-tRNA Ligase/chemistry/*metabolism Base Sequence Crystallization Escherichia coli/*enzymology/genetics Glutamate-tRNA Ligase/chemistry/*metabolism Kinetics Molecular Sequence Data Molecular Structure Mutation Nucleic Acid Conformation RNA, Asp/chemistry/*metabolism RNA, ERIANI, FLORENTZ, FRUGIER, Gln/chemistry/*metabolism Saccharomyces cerevisiae/*enzymology/genetics Support, Non-U.S. Gov't Support, P.H.S., Transfer, U.S. Gov't, Unité ARN
@article{,
title = {Identity switches between tRNAs aminoacylated by class I glutaminyl- and class II aspartyl-tRNA synthetases},
author = {M Frugier and D Soll and R Giege and C Florentz},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8060999},
isbn = {8060999},
year = {1994},
date = {1994-01-01},
journal = {Biochemistry},
volume = {33},
number = {33},
pages = {9912-9921},
abstract = {High-resolution X-ray structures for the tRNA/aminoacyl-tRNA synthetase complexes between Escherichia coli tRNAGln/GlnRS and yeast tRNAAsp/AspRS have been determined. Positive identity nucleotides that direct aminoacylation specificity have been defined in both cases; E. coli tRNAGln identity is governed by 10 elements scattered in the tRNA structure, while specific aminoacylation of yeast tRNAAsp is dependent on 5 positions. Both identity sets are partially overlapping and share 3 nucleotides. Interestingly, the two enzymes belong to two different classes described for aminoacyl-tRNA synthetases. The class I glutaminyl-tRNA synthetase and the class II aspartyl-tRNA synthetase recognize their cognate tRNA from opposite sides. Mutants derived from glutamine and aspartate tRNAs have been created by progressively introducing identity elements from one tRNA into the other one. Glutaminylation and aspartylation assays of the transplanted tRNAs show that identity nucleotides from a tRNA originally aminoacylated by a synthetase from one class are still recognized if they are presented to the enzyme in a structural framework corresponding to a tRNA aminoacylated by a synthetase belonging to the other class. The simple transplantation of the glutamine identity set into tRNAAsp is sufficient to obtain glutaminylatable tRNA, but additional subtle features seem to be important for the complete conversion of tRNAGln in an aspartylatable substrate. This study defines C38 in yeast tRNAAsp as a new identity nucleotide for aspartylation. We show also in this paper that, during the complex formation, aminoacyl-tRNA synthetases are at least partially responsible for conformational changes which involve structural constraints in tRNA molecules.},
note = {0006-2960
Journal Article},
keywords = {Acylation Aspartate-tRNA Ligase/chemistry/*metabolism Base Sequence Crystallization Escherichia coli/*enzymology/genetics Glutamate-tRNA Ligase/chemistry/*metabolism Kinetics Molecular Sequence Data Molecular Structure Mutation Nucleic Acid Conformation RNA, Asp/chemistry/*metabolism RNA, ERIANI, FLORENTZ, FRUGIER, Gln/chemistry/*metabolism Saccharomyces cerevisiae/*enzymology/genetics Support, Non-U.S. Gov't Support, P.H.S., Transfer, U.S. Gov't, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Frugier M, Florentz C, Hosseini M W, Lehn J M, Giege R
Synthetic polyamines stimulate in vitro transcription by T7 RNA polymerase Article de journal
Dans: Nucleic Acids Res, vol. 22, no. 14, p. 2784-2790, 1994, ISBN: 8052534, (0305-1048 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: 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}
}
Frugier M, Florentz C, Giege R
Efficient aminoacylation of resected RNA helices by class II aspartyl-tRNA synthetase dependent on a single nucleotide Article de journal
Dans: EMBO J, vol. 13, no. 9, p. 2218-2226, 1994, ISBN: 8187774, (0261-4189 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Acylation Anticodon Aspartate-tRNA Ligase/*metabolism Aspartic Acid/metabolism Base Sequence Evolution Molecular Sequence Data Nucleic Acid Conformation RNA, ERIANI, FLORENTZ, FRUGIER, Fungal/chemistry/*metabolism Substrate Specificity Support, Non-U.S. Gov't, Unité ARN
@article{,
title = {Efficient aminoacylation of resected RNA helices by class II aspartyl-tRNA synthetase dependent on a single nucleotide},
author = {M Frugier and C Florentz and R Giege},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8187774},
isbn = {8187774},
year = {1994},
date = {1994-01-01},
journal = {EMBO J},
volume = {13},
number = {9},
pages = {2218-2226},
abstract = {We show here that small RNA helices which recapitulate part or all of the acceptor stem of yeast aspartate tRNA are efficiently aminoacylated by cognate class II aspartyl-tRNA synthetase. Aminoacylation is strongly dependent on the presence of the single-stranded G73 'discriminator' identity nucleotide and is essentially insensitive to the sequence of the helical region. Substrates which contain as few as 3 bp fused to G73CCAOH are aspartylated. Their charging is insensitive to the sequence of the loop closing the short helical domains. Aminoacylation of the aspartate mini-helix is not stimulated by a hairpin helix mimicking the anticodon domain and containing the three major anticodon identity nucleotides. A thermodynamic analysis demonstrates that enzyme interactions with G73 in the resected RNA substrates and in the whole tRNA are the same. Thus, if the resected RNA molecules resemble in some way the earliest substrates for aminoacylation with aspartate, then the contemporary tRNA(Asp) has quantitatively retained the influence of the major signal for aminoacylation in these substrates.},
note = {0261-4189
Journal Article},
keywords = {Acylation Anticodon Aspartate-tRNA Ligase/*metabolism Aspartic Acid/metabolism Base Sequence Evolution Molecular Sequence Data Nucleic Acid Conformation RNA, ERIANI, FLORENTZ, FRUGIER, Fungal/chemistry/*metabolism Substrate Specificity Support, Non-U.S. Gov't, Unité ARN},
pubstate = {published},
tppubtype = {article}
}