Publications
2004
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}
}
2003
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}
}
2000
Helm M, Brule H, Friede D, Giege R, Putz D, Florentz C
Search for characteristic structural features of mammalian mitochondrial tRNAs Article de journal
Dans: RNA, vol. 6, no. 10, p. 1356-1379, 2000, ISBN: 11073213, (1355-8382 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Acylation Animals Base Pairing Base Sequence *Computational Biology Escherichia coli/genetics Genome Human Molecular Sequence Data Multigene Family *Nucleic Acid Conformation RNA/*chemistry/genetics RNA Stability RNA, Amino Acid-Specific/*chemistry/genetics Regulatory Sequences, FLORENTZ, Non-U.S. Gov't Variation (Genetics), Nucleic Acid/genetics Support, Transfer, Unité ARN
@article{,
title = {Search for characteristic structural features of mammalian mitochondrial tRNAs},
author = {M Helm and H Brule and D Friede and R Giege and D Putz and C Florentz},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11073213},
isbn = {11073213},
year = {2000},
date = {2000-01-01},
journal = {RNA},
volume = {6},
number = {10},
pages = {1356-1379},
abstract = {A number of mitochondrial (mt) tRNAs have strong structural deviations from the classical tRNA cloverleaf secondary structure and from the conventional L-shaped tertiary structure. As a consequence, there is a general trend to consider all mitochondrial tRNAs as "bizarre" tRNAs. Here, a large sequence comparison of the 22 tRNA genes within 31 fully sequenced mammalian mt genomes has been performed to define the structural characteristics of this specific group of tRNAs. Vertical alignments define the degree of conservation/variability of primary sequences and secondary structures and search for potential tertiary interactions within each of the 22 families. Further horizontal alignments ascertain that, with the exception of serine-specific tRNAs, mammalian mt tRNAs do fold into cloverleaf structures with mostly classical features. However, deviations exist and concern large variations in size of the D- and T-loops. The predominant absence of the conserved nucleotides G18G19 and T54T55C56, respectively in these loops, suggests that classical tertiary interactions between both domains do not take place. Classification of the tRNA sequences according to their genomic origin (G-rich or G-poor DNA strand) highlight specific features such as richness/poorness in mismatches or G-T pairs in stems and extremely low G-content or C-content in the D- and T-loops. The resulting 22 "typical" mammalian mitochondrial sequences built up a phylogenetic basis for experimental structural and functional investigations. Moreover, they are expected to help in the evaluation of the possible impacts of those point mutations detected in human mitochondrial tRNA genes and correlated with pathologies.},
note = {1355-8382
Journal Article},
keywords = {Acylation Animals Base Pairing Base Sequence *Computational Biology Escherichia coli/genetics Genome Human Molecular Sequence Data Multigene Family *Nucleic Acid Conformation RNA/*chemistry/genetics RNA Stability RNA, Amino Acid-Specific/*chemistry/genetics Regulatory Sequences, FLORENTZ, Non-U.S. Gov't Variation (Genetics), Nucleic Acid/genetics Support, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
1997
Massire C, Jaeger L, Westhof E
Phylogenetic evidence for a new tertiary interaction in bacterial RNase P RNAs Article de journal
Dans: RNA, vol. 3, no. 6, p. 553-556, 1997, ISBN: 9174090, (1355-8382 Letter).
Liens | BibTeX | Étiquettes: Bacterial/*chemistry RNA, Base Sequence Comparative Study Databases, Catalytic/*chemistry Ribonuclease P Sequence Homology, Factual Endoribonucleases/*chemistry Molecular Sequence Data *Nucleic Acid Conformation Phylogeny RNA, Non-U.S. Gov't Variation (Genetics), Nucleic Acid Support, Unité ARN
@article{,
title = {Phylogenetic evidence for a new tertiary interaction in bacterial RNase P RNAs},
author = {C Massire and L Jaeger and E Westhof},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=9174090},
isbn = {9174090},
year = {1997},
date = {1997-01-01},
journal = {RNA},
volume = {3},
number = {6},
pages = {553-556},
note = {1355-8382
Letter},
keywords = {Bacterial/*chemistry RNA, Base Sequence Comparative Study Databases, Catalytic/*chemistry Ribonuclease P Sequence Homology, Factual Endoribonucleases/*chemistry Molecular Sequence Data *Nucleic Acid Conformation Phylogeny RNA, Non-U.S. Gov't Variation (Genetics), Nucleic Acid Support, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
1996
Aphasizhev R, Senger B, Rengers J U, Sprinzl M, Walter P, Nussbaum G, Fasiolo F
Conservation in evolution for a small monomeric phenylalanyl-tRNA synthetase of the tRNA(Phe) recognition nucleotides and initial aminoacylation site Article de journal
Dans: Biochemistry, vol. 35, no. 1, p. 117-123, 1996, ISBN: 8555164, (0006-2960 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Non-U.S. Gov't Variation (Genetics), Nucleic Acid Substrate Specificity Support, Phe/*chemistry/*metabolism Saccharomyces cerevisiae/enzymology/genetics Sequence Homology, Transfer, Unité ARN
@article{,
title = {Conservation in evolution for a small monomeric phenylalanyl-tRNA synthetase of the tRNA(Phe) recognition nucleotides and initial aminoacylation site},
author = {R Aphasizhev and B Senger and J U Rengers and M Sprinzl and P Walter and G Nussbaum and F Fasiolo},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8555164},
isbn = {8555164},
year = {1996},
date = {1996-01-01},
journal = {Biochemistry},
volume = {35},
number = {1},
pages = {117-123},
abstract = {We previously showed that yeast mitochondrial phenylalanyl-tRNA synthetase (MSF protein) is evolutionarily distant to the cytoplasmic counterpart based on a high degree of divergence in protein sequence, molecular mass, and quaternary structure. Using yeast cytoplasmic tRNA(Phe) which is efficiently aminoacylated by MSF protein, we report here the tRNA(Phe) primary site of aminoacylation and the identity determinants for MSF protein. As for the cytoplasmic phenylalanyl-tRNA synthetase (Sampson, J. R., Di Renzo, A. B., Behlen, L. S., & Uhlenbeck, O. C. (1989) Science 243, 1363-1366), MSF protein recognizes nucleotides from the anticodon and the acceptor end including base A73 and, as shown here, adjacent G1-C72 base pair or at least C72 base. This indicates that the way of tRNA(Phe) binding for the two phenylalanine enzymes is conserved in evolution. However, tRNA(Phe) tertiary structure seems more critical for the interaction with the cytoplasmic enzyme than with MSF protein, and unlike cytoplasmic phenylalanyl-tRNA synthetase, the small size of the monomeric MSF protein probably does not allow contacts with residue 20 at the top corner of the L molecule. We also show that MSF protein preferentially aminoacylates the terminal 2'-OH group of tRNA(Phe) but with a catalytic efficiency for tRNA(Phe)-CC-3'-deoxyadenosine reduced 100-fold from that of native tRNA(Phe), suggesting a role of the terminal 3'-OH in catalysis. The loss is only 1.5-fold when tRNA(Phe)-CC-3'-deoxyadenosine is aminoacylated by yeast cytoplasmic PheRS (Sprinzl, M., & Cramer, F. (1973) Nature 245, 3-5), indicating mechanistic differences between the two PheRS's active sites for the amino acid transfer step.},
note = {0006-2960
Journal Article},
keywords = {Non-U.S. Gov't Variation (Genetics), Nucleic Acid Substrate Specificity Support, Phe/*chemistry/*metabolism Saccharomyces cerevisiae/enzymology/genetics Sequence Homology, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}