Publications
2004
Sissler M, Helm M, Frugier M, Giege R, Florentz C
Aminoacylation properties of pathology-related human mitochondrial tRNA(Lys) variants Journal Article
In: RNA, vol. 10, no. 5, pp. 841-853, 2004, ISBN: 15100439, (1355-8382 Journal Article).
Abstract | Links | BibTeX | Tags: 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}
}
2000
Wientges J, Putz J, Giege R, Florentz C, Schwienhorst A
Selection of viral RNA-derived tRNA-like structures with improved valylation activities Journal Article
In: Biochemistry, vol. 39, no. 20, pp. 6207-6218, 2000, ISBN: 10821696, (0006-2960 Journal Article).
Abstract | Links | BibTeX | Tags: 3' Untranslated Regions Acylation Anticodon/chemistry Base Sequence Cloning, FLORENTZ, Molecular Gene Library Kinetics Molecular Sequence Data Nucleic Acid Conformation Oligonucleotides/chemistry RNA, Non-U.S. Gov't Tymovirus/enzymology/genetics Valine-tRNA Ligase/chemistry Variation (Genetics), RNA Support, Transfer, Unité ARN, Val/*chemistry RNA, Viral/*chemistry Sequence Analysis
@article{,
title = {Selection of viral RNA-derived tRNA-like structures with improved valylation activities},
author = {J Wientges and J Putz and R Giege and C Florentz and A Schwienhorst},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=10821696},
isbn = {10821696},
year = {2000},
date = {2000-01-01},
journal = {Biochemistry},
volume = {39},
number = {20},
pages = {6207-6218},
abstract = {The tRNA-like structure (TLS) of turnip yellow mosaic virus (TYMV) RNA was previously shown to be efficiently charged by yeast valyl-tRNA synthetase (ValRS). This RNA has a noncanonical structure at its 3'-terminus but mimics a tRNA L-shaped fold, including an anticodon loop containing the major identity nucleotides for valylation, and a pseudoknotted amino acid accepting domain. Here we describe an in vitro selection experiment aimed (i) to verify the completeness of the valine identity set, (ii) to elucidate the impact of the pseudoknot on valylation, and (iii) to investigate whether functional communication exists between the two distal anticodon and amino acid accepting domains. Valylatable variants were selected from a pool of 2 x 10(13) RNA molecules derived from the TYMV TLS randomized in the anticodon loop nucleotides and in the length (1-6 nucleotides) and sequence of the pseudoknot loop L1. After nine rounds of selection by aminoacylation, 42 have been isolated. Among them, 17 RNAs could be efficiently charged by yeast ValRS. Their sequence revealed strong conservation of the second and the third anticodon triplet positions (A(56), C(55)) and the very 3'-end loop nucleotide C(53). A large variability of the other nucleotides of the loop was observed and no wild-type sequence was recovered. The selected molecules presented pseudoknot domains with loop L1 varying in size from 3-6 nucleotides and some sequence conservation, but did neither reveal the wild-type combination. All selected variants are 5-50 times more efficiently valylated than the wild-type TLS, suggesting that the natural viral sequence has emerged from a combination of evolutionary pressures among which aminoacylation was not predominant. This is in line with the role of the TLS in viral replication.},
note = {0006-2960
Journal Article},
keywords = {3' Untranslated Regions Acylation Anticodon/chemistry Base Sequence Cloning, FLORENTZ, Molecular Gene Library Kinetics Molecular Sequence Data Nucleic Acid Conformation Oligonucleotides/chemistry RNA, Non-U.S. Gov't Tymovirus/enzymology/genetics Valine-tRNA Ligase/chemistry Variation (Genetics), RNA Support, Transfer, Unité ARN, Val/*chemistry RNA, Viral/*chemistry Sequence Analysis},
pubstate = {published},
tppubtype = {article}
}
1997
Friant S, Heyman T, Poch O, Wilhelm M, Wilhelm F X
In: Yeast, vol. 13, no. 7, pp. 639-645, 1997, ISBN: 9200813, (0749-503x Journal Article).
Abstract | Links | BibTeX | Tags: Amino Acid Sequence Base Sequence DNA Transposable Elements/*genetics Molecular Sequence Data Molecular Structure RNA, Fungal/genetics RNA, Met/*chemistry/*genetics Sequence Alignment *Sequence Analysis, Non-U.S. Gov't Yeasts/*genetics, RNA Support, Transfer, Unité ARN
@article{,
title = {Sequence comparison of the Ty1 and Ty2 elements of the yeast genome supports the structural model of the tRNAiMet-Ty1 RNA reverse transcription initiation complex},
author = {S Friant and T Heyman and O Poch and M Wilhelm and F X Wilhelm},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9200813},
isbn = {9200813},
year = {1997},
date = {1997-01-01},
journal = {Yeast},
volume = {13},
number = {7},
pages = {639-645},
abstract = {In the reverse transcription initiation complex of the yeast Ty1 retrotransposon, interaction between the template RNA and primer tRNAiMet is not limited to base pairing of the primer binding site (PBS) with ten nucleotides at the 3' end of tRNAiMet, but three regions named boxes O, 1 and 2.1 interact with the T and D stems and loops of tRNAiMet. Sequence comparison of 33 Ty1 elements and 13 closely related Ty2 elements found in the yeast genome shows that the nucleotide sequence of all elements is highly conserved in the region spanning the PBS and the three boxes. Since the domain of the template RNA encodes a portion of protein TyA, we have calculated its amino acid profile and its nucleotide profile to evaluate the role played by nucleotide sequence conservation in the selection for TyA function and in the maintenance of base pairing interactions for the priming function of Ty1 RNA. Our results show that the nucleotide sequence conservation of Ty1 RNA is constrained not only by selection for Ty1 function but also by maintenance of a given nucleotide sequence able to base pair with the tRNAiMet in the primer-template initiation complex.},
note = {0749-503x
Journal Article},
keywords = {Amino Acid Sequence Base Sequence DNA Transposable Elements/*genetics Molecular Sequence Data Molecular Structure RNA, Fungal/genetics RNA, Met/*chemistry/*genetics Sequence Alignment *Sequence Analysis, Non-U.S. Gov't Yeasts/*genetics, RNA Support, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}