Cavarelli J, Eriani G, Rees B, Ruff M, Boeglin M, Mitschler A, Martin F, Gangloff J, Thierry J C, Moras D
The active site of yeast aspartyl-tRNA synthetase: structural and functional aspects of the aminoacylation reaction Article de journal
Dans: EMBO J, vol. 13, no. 2, p. 327-337, 1994, ISBN: 8313877, (0261-4189 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Acylation Adenosine Triphosphate/metabolism Amino Acid Sequence Animals Aspartate-tRNA Ligase/chemistry/genetics/*metabolism Binding Sites Computer Graphics Human Molecular Sequence Data Mutagenesis, Amino Acid Structure-Activity Relationship Support, ERIANI, Non-U.S. Gov't, Site-Directed Saccharomyces cerevisiae/*enzymology Sequence Homology, Unité ARN
@article{,
title = {The active site of yeast aspartyl-tRNA synthetase: structural and functional aspects of the aminoacylation reaction},
author = {J Cavarelli and G Eriani and B Rees and M Ruff and M Boeglin and A Mitschler and F Martin and J Gangloff and J C Thierry and D Moras},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8313877},
isbn = {8313877},
year = {1994},
date = {1994-01-01},
journal = {EMBO J},
volume = {13},
number = {2},
pages = {327-337},
abstract = {The crystal structures of the various complexes formed by yeast aspartyl-tRNA synthetase (AspRS) and its substrates provide snapshots of the active site corresponding to different steps of the aminoacylation reaction. Native crystals of the binary complex tRNA-AspRS were soaked in solutions containing the two other substrates, ATP (or its analog AMPPcP) and aspartic acid. When all substrates are present in the crystal, this leads to the formation of the aspartyl-adenylate and/or the aspartyl-tRNA. A class II-specific pathway for the aminoacylation reaction is proposed which explains the known functional differences between the two classes while preserving a common framework. Extended signature sequences characteristic of class II aaRS (motifs 2 and 3) constitute the basic functional unit. The ATP molecule adopts a bent conformation, stabilized by the invariant Arg531 of motif 3 and a magnesium ion coordinated to the pyrophosphate group and to two class-invariant acidic residues. The aspartic acid substrate is positioned by a class II invariant acidic residue, Asp342, interacting with the amino group and by amino acids conserved in the aspartyl synthetase family. The amino acids in contact with the substrates have been probed by site-directed mutagenesis for their functional implication.},
note = {0261-4189
Journal Article},
keywords = {Acylation Adenosine Triphosphate/metabolism Amino Acid Sequence Animals Aspartate-tRNA Ligase/chemistry/genetics/*metabolism Binding Sites Computer Graphics Human Molecular Sequence Data Mutagenesis, Amino Acid Structure-Activity Relationship Support, ERIANI, Non-U.S. Gov't, Site-Directed Saccharomyces cerevisiae/*enzymology Sequence Homology, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Frugier M, Florentz C, Schimmel P, Giege R
Triple aminoacylation specificity of a chimerized transfer RNA Article de journal
Dans: Biochemistry, vol. 32, no. 50, p. 14053-14061, 1993, ISBN: 8268184, (0006-2960 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Acylation Alanine/metabolism Base Sequence Chimera Escherichia coli/genetics Molecular Sequence Data Mutation Nucleic Acid Conformation Phenylalanine/metabolism RNA, Asp/chemistry/genetics/*metabolism Saccharomyces cerevisiae/genetics Support, ERIANI, FLORENTZ, FRUGIER, Non-U.S. Gov't Support, P.H.S. Valine/metabolism, Transfer, U.S. Gov't, Unité ARN
@article{,
title = {Triple aminoacylation specificity of a chimerized transfer RNA},
author = {M Frugier and C Florentz and P Schimmel and R Giege},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8268184},
isbn = {8268184},
year = {1993},
date = {1993-01-01},
journal = {Biochemistry},
volume = {32},
number = {50},
pages = {14053-14061},
abstract = {We report here the rational design and construction of a chimerized transfer RNA with tripartite aminoacylation specificity. A yeast aspartic acid specific tRNA was transformed into a highly efficient acceptor of alanine and phenylalanine and a moderate acceptor of valine. The transformation was guided by available knowledge of the requirements for aminoacylation by each of the three amino acids and was achieved by iterative changes in the local sequence context and the structural framework of the variable loop and the two variable regions of the dihydrouridine loop. The changes introduced to confer efficient acceptance of the three amino acids eliminate aminoacylation with aspartate. The interplay of determinants and antideterminants for different specific aminoacylations, and the constraints imposed by the structural framework, suggest that a tRNA with an appreciable capacity for more than three efficient aminoacylations may be inherently difficult to achieve.},
note = {0006-2960
Journal Article},
keywords = {Acylation Alanine/metabolism Base Sequence Chimera Escherichia coli/genetics Molecular Sequence Data Mutation Nucleic Acid Conformation Phenylalanine/metabolism RNA, Asp/chemistry/genetics/*metabolism Saccharomyces cerevisiae/genetics Support, ERIANI, FLORENTZ, FRUGIER, Non-U.S. Gov't Support, P.H.S. Valine/metabolism, Transfer, U.S. Gov't, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Roth A, Eriani G, Dirheimer G, Gangloff J
Kinetic properties of pure overproduced Bacillus subtilis phenylalanyl-tRNA synthetase do not favour its in vivo inhibition by ochratoxin A Article de journal
Dans: FEBS Lett, vol. 326, no. 1-3, p. 87-91, 1993, ISBN: 8325392, (0014-5793 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Bacillus subtilis/drug effects/*enzymology Binding, Bacterial, Competitive Escherichia coli/enzymology/genetics Kinetics Macromolecular Systems Ochratoxins/*pharmacology Phenylalanine/metabolism Phenylalanine-tRNA Ligase/antagonists & inhibitors/*metabolism Support, ERIANI, Non-U.S. Gov't Transformation, Unité ARN
@article{,
title = {Kinetic properties of pure overproduced Bacillus subtilis phenylalanyl-tRNA synthetase do not favour its in vivo inhibition by ochratoxin A},
author = {A Roth and G Eriani and G Dirheimer and J Gangloff},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8325392},
isbn = {8325392},
year = {1993},
date = {1993-01-01},
journal = {FEBS Lett},
volume = {326},
number = {1-3},
pages = {87-91},
abstract = {Ochratoxine A (OTA) inhibits growth of Bacillus subtilis at pHs below 7. Since OTA is a phenylalanine analogue, this effect could be due to inhibition of phenylalanine-tRNA synthetase (PheRS) by competition of this mycotoxin with the amino acid. Homogeneous PheRS was purified from Bacillus subtilis and from E. coli transformed with the PheRS gene. The latter produced about 40 times more PheRS than B. subtilis. The Km and Ki values of PheRS, respectively, for phenylalanine and OTA were measured and their concentrations within the cell determined. It appears that the concentration of OTA in the cell, in spite of a 25-fold accumulation, remained too low to significantly compete with phenylalanine. This does not suggest PheRS to be the target of OTA in cell growth and protein synthesis inhibition in Bacillus subtilis. It was also shown that the 2-3-fold increase of PheRS in OTA-treated cells is not due to phenylalanine-controlled attenuation regulation.},
note = {0014-5793
Journal Article},
keywords = {Bacillus subtilis/drug effects/*enzymology Binding, Bacterial, Competitive Escherichia coli/enzymology/genetics Kinetics Macromolecular Systems Ochratoxins/*pharmacology Phenylalanine/metabolism Phenylalanine-tRNA Ligase/antagonists & inhibitors/*metabolism Support, ERIANI, Non-U.S. Gov't Transformation, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Oguiza J A, Malumbres M, Eriani G, Pisabarro A, Mateos L M, Martin F, Martin J F
Dans: J Bacteriol, vol. 175, no. 22, p. 7356-7362, 1993, ISBN: 8226683, (0021-9193 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Amino Acid Support, Amino Acid Sequence Amino Acyl-tRNA Ligases/genetics Arginine/*pharmacology Arginine-tRNA Ligase/biosynthesis/*genetics Bacteria/enzymology Brevibacterium/*enzymology/*genetics Carboxy-Lyases/*genetics Cloning, Bacterial Molecular Sequence Data Molecular Weight *Multigene Family Plasmids Restriction Mapping Sequence Homology, Bacterial/*drug effects Gene Expression Regulation, Enzymologic/drug effects *Genes, ERIANI, Molecular Comparative Study Escherichia coli/genetics/growth & development Fungi/enzymology Gene Expression Regulation, Non-U.S. Gov't, Structural, Unité ARN
@article{,
title = {A gene encoding arginyl-tRNA synthetase is located in the upstream region of the lysA gene in Brevibacterium lactofermentum: regulation of argS-lysA cluster expression by arginine},
author = {J A Oguiza and M Malumbres and G Eriani and A Pisabarro and L M Mateos and F Martin and J F Martin},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8226683},
isbn = {8226683},
year = {1993},
date = {1993-01-01},
journal = {J Bacteriol},
volume = {175},
number = {22},
pages = {7356-7362},
abstract = {The Brevibacterium lactofermentum argS gene, which encodes an arginyl-tRNA synthetase, was identified in the upstream region of the lysA gene. The cloned gene was sequenced; it encodes a 550-amino-acid protein with an M(r) of 59,797. The deduced amino acid sequence showed 28% identical and 49% similar residues when compared with the sequence of the Escherichia coli arginyl-tRNA synthetase. The B. lactofermentum enzyme showed the highly conserved motifs of class I aminoacyl-tRNA synthetases. Expression of the argS gene in B. lactofermentum and E. coli resulted in an increase in aminoacyl-tRNA synthetase activity, correlated with the presence in sodium dodecyl sulfate-polyacrylamide gels of a clear protein band that corresponds to this enzyme. One single transcript of about 3,000 nucleotides and corresponding to the B. lactofermentum argS-lysA operon was identified. The transcription of these genes is repressed by lysine and induced by arginine, showing an interesting pattern of biosynthetic interlock between the pathways of both amino acids in corynebacteria.},
note = {0021-9193
Journal Article},
keywords = {Amino Acid Support, Amino Acid Sequence Amino Acyl-tRNA Ligases/genetics Arginine/*pharmacology Arginine-tRNA Ligase/biosynthesis/*genetics Bacteria/enzymology Brevibacterium/*enzymology/*genetics Carboxy-Lyases/*genetics Cloning, Bacterial Molecular Sequence Data Molecular Weight *Multigene Family Plasmids Restriction Mapping Sequence Homology, Bacterial/*drug effects Gene Expression Regulation, Enzymologic/drug effects *Genes, ERIANI, Molecular Comparative Study Escherichia coli/genetics/growth & development Fungi/enzymology Gene Expression Regulation, Non-U.S. Gov't, Structural, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Martin F, Eriani G, Eiler S, Moras D, Dirheimer G, Gangloff J
Overproduction and purification of native and queuine-lacking Escherichia coli tRNA(Asp). Role of the wobble base in tRNA(Asp) acylation Article de journal
Dans: J Mol Biol, vol. 234, no. 4, p. 965-974, 1993, ISBN: 8263943, (0022-2836 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: *Amino Acid Activation Anticodon Aspartate-tRNA Ligase/metabolism Base Composition Base Sequence Cloning, Asp/chemistry/*metabolism Saccharomyces cerevisiae/metabolism Structure-Activity Relationship Support, ERIANI, Molecular Comparative Study Crystallography, Non-U.S. Gov't, Transfer, Unité ARN, X-Ray Escherichia coli/*metabolism Guanine/*analogs & derivatives/chemistry Molecular Sequence Data Nucleic Acid Conformation RNA
@article{,
title = {Overproduction and purification of native and queuine-lacking Escherichia coli tRNA(Asp). Role of the wobble base in tRNA(Asp) acylation},
author = {F Martin and G Eriani and S Eiler and D Moras and G Dirheimer and J Gangloff},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8263943},
isbn = {8263943},
year = {1993},
date = {1993-01-01},
journal = {J Mol Biol},
volume = {234},
number = {4},
pages = {965-974},
abstract = {Escherichia coli tRNA(Asp) was overproduced in E. coli up to 15-fold from a synthetic tRNA(Asp) gene placed in a plasmid under the dependence of an isopropyl-beta,D-thiogalactopyranoside-inducible promoter. Purification to nearly homogeneity (95%) was achieved after two HPLC DEAE-cellulose columns. E. coli tRNA(Asp)[G34] (having guanine instead of queuine at position 34) was obtained by the same procedure except that it was overproduced in a strain lacking the enzyme responsible for queuine modification. Nucleoside analysis showed that, except for the replacement of Q34 by G34 in mutant-derived tRNA(Asp), the base modification levels of both tRNAs are the same as those in wild-type E. coli tRNA(Asp). Kinetic properties of tRNA(Asp)[Q34] and [G34] with yeast AspRS compared to those in the homologous reactions in yeast and E. coli clearly indicate that the major identity elements are the same in both organisms: the conserved discriminant base and the anticodon triplet. In connection with this, we explored by site-directed mutagenesis the functional role of the interactions which, as revealed by the crystallographic structure, occur between the wobble base of yeast tRNA(Asp) and two residues of yeast AspRS. Their absence strongly affected aspartylation and the kd of tRNA(Asp). Each contact individually restores almost completely the wild-type acylation properties of the enzyme; thus, wobble base recognition in yeast appears to be more protected against mutational events than in E. coli, where only one contact is thought to occur at position 34.},
note = {0022-2836
Journal Article},
keywords = {*Amino Acid Activation Anticodon Aspartate-tRNA Ligase/metabolism Base Composition Base Sequence Cloning, Asp/chemistry/*metabolism Saccharomyces cerevisiae/metabolism Structure-Activity Relationship Support, ERIANI, Molecular Comparative Study Crystallography, Non-U.S. Gov't, Transfer, Unité ARN, X-Ray Escherichia coli/*metabolism Guanine/*analogs & derivatives/chemistry Molecular Sequence Data Nucleic Acid Conformation RNA},
pubstate = {published},
tppubtype = {article}
}
Eriani G, Cavarelli J, Martin F, Dirheimer G, Moras D, Gangloff J
Role of dimerization in yeast aspartyl-tRNA synthetase and importance of the class II invariant proline Article de journal
Dans: Proc Natl Acad Sci U S A, vol. 90, no. 22, p. 10816-10820, 1993, ISBN: 8248175, (0027-8424 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Asp/metabolism Saccharomyces cerevisiae/chemistry Structure-Activity Relationship Support, Aspartate-tRNA Ligase/*chemistry Fungal Proteins/chemistry Kinetics Macromolecular Systems Models, ERIANI, Molecular Mutagenesis, Non-U.S. Gov't, Site-Directed Proline/chemistry Protein Binding Protein Conformation RNA, Transfer, Unité ARN
@article{,
title = {Role of dimerization in yeast aspartyl-tRNA synthetase and importance of the class II invariant proline},
author = {G Eriani and J Cavarelli and F Martin and G Dirheimer and D Moras and J Gangloff},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8248175},
isbn = {8248175},
year = {1993},
date = {1993-01-01},
journal = {Proc Natl Acad Sci U S A},
volume = {90},
number = {22},
pages = {10816-10820},
abstract = {Cytoplasmic aspartyl-tRNA synthetase (AspRS; EC 6.1.1.12) from yeast is, as are most class II synthetases, an alpha 2 dimer. The only invariant amino acid in signature motif 1 of this class is Pro-273; this residue is located at the dimer interface. To understand the role of Pro-273 in the conserved dimeric configuration, we tested the effect of a Pro-273-->Gly (P273G) substitution on the catalytic properties of homo- and heterodimeric AspRS. Heterodimers of AspRS were produced in vivo by overexpression of their respective subunit variants from plasmid-encoded genes and purified to homogeneity in one HPLC step. The homodimer containing the P273G shows an 80% inactivation of the enzyme and an affinity decrease for its cognate tRNA(Asp) of one order of magnitude. The P273G-mutated subunit recovered wild-type enzymatic properties when associated with a native subunit or a monomer otherwise inactivated having an intact dimeric interface domain. These results, which can be explained by the crystal structure of the native enzyme complexed with its substrates, confirm the structural importance of Pro-273 for dimerization and clearly establish the functional interdependence of the AspRS subunits. More generally, the dimeric conformation may be a structural prerequisite for the activity of mononucleotide binding sites constructed from antiparallel beta strands.},
note = {0027-8424
Journal Article},
keywords = {Asp/metabolism Saccharomyces cerevisiae/chemistry Structure-Activity Relationship Support, Aspartate-tRNA Ligase/*chemistry Fungal Proteins/chemistry Kinetics Macromolecular Systems Models, ERIANI, Molecular Mutagenesis, Non-U.S. Gov't, Site-Directed Proline/chemistry Protein Binding Protein Conformation RNA, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
Frugier M, Florentz C, Giege R
Anticodon-independent aminoacylation of an RNA minihelix with valine Article de journal
Dans: Proc Natl Acad Sci U S A, vol. 89, no. 9, p. 3990-3994, 1992, ISBN: 1570324, (0027-8424 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: *Amino Acid Activation Anticodon Base Sequence Hydrogen Bonding In Vitro Molecular Sequence Data RNA, ERIANI, FLORENTZ, FRUGIER, Non-U.S. Gov't Valine-tRNA Ligase/*metabolism, Transfer, Unité ARN, Val/chemistry/*metabolism Saccharomyces cerevisiae Structure-Activity Relationship Support
@article{,
title = {Anticodon-independent aminoacylation of an RNA minihelix with valine},
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=1570324},
isbn = {1570324},
year = {1992},
date = {1992-01-01},
journal = {Proc Natl Acad Sci U S A},
volume = {89},
number = {9},
pages = {3990-3994},
abstract = {Minihelices mimicking the amino acid acceptor and anticodon branches of yeast tRNA(Val) have been synthesized by in vitro transcription of synthetic templates. It is shown that a minihelix corresponding to the amino acid acceptor branch and containing solely a valine-specific identity nucleotide can be aminoacylated by yeast valyl-tRNA synthetase. Its charging ability is lost after mutating this nucleotide. This ability is stimulated somewhat by the addition of a second hairpin helix that mimicks the anticodon arm, which suggests that information originating from the anticodon stem-loop can be transmitted to the active site of the enzyme by the core of the protein.},
note = {0027-8424
Journal Article},
keywords = {*Amino Acid Activation Anticodon Base Sequence Hydrogen Bonding In Vitro Molecular Sequence Data RNA, ERIANI, FLORENTZ, FRUGIER, Non-U.S. Gov't Valine-tRNA Ligase/*metabolism, Transfer, Unité ARN, Val/chemistry/*metabolism Saccharomyces cerevisiae Structure-Activity Relationship Support},
pubstate = {published},
tppubtype = {article}
}
Eiler S, Boeglin M, Martin F, Eriani G, Gangloff J, Thierry J C, Moras D
Crystallization of aspartyl-tRNA synthetase-tRNA(Asp) complex from Escherichia coli and first crystallographic results Article de journal
Dans: J Mol Biol, vol. 224, no. 4, p. 1171-1173, 1992, ISBN: 1569573, (0022-2836 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Asp/*ultrastructure X-Ray Diffraction, Aspartate-tRNA Ligase/*ultrastructure Crystallography Escherichia coli/enzymology RNA, ERIANI, Transfer, Unité ARN
@article{,
title = {Crystallization of aspartyl-tRNA synthetase-tRNA(Asp) complex from Escherichia coli and first crystallographic results},
author = {S Eiler and M Boeglin and F Martin and G Eriani and J Gangloff and J C Thierry and D Moras},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=1569573},
isbn = {1569573},
year = {1992},
date = {1992-01-01},
journal = {J Mol Biol},
volume = {224},
number = {4},
pages = {1171-1173},
abstract = {Crystals of the dimeric aspartyl-tRNA synthetase from Escherichia coli (molecular mass 132,000 Da) complexed with its cognate tRNA (molecular mass 25,000 Da) have been grown using ammonium sulfate as precipitant. The crystals belong to the orthorhombic space group C222(1) with unit cell parameters a = 102.75 A},
note = {0022-2836
Journal Article},
keywords = {Asp/*ultrastructure X-Ray Diffraction, Aspartate-tRNA Ligase/*ultrastructure Crystallography Escherichia coli/enzymology RNA, ERIANI, Transfer, Unité ARN},
pubstate = {published},
tppubtype = {article}
}