title: Structural and kinetic bases for the recognition of tRNATyr by tyrosyl-tRNA synthetase.
abstract: The aminoacylation of transfer RNA is a key step of translation since it relates amino acids to anticodons. To understand how the tyrosyl-tRNA synthetase (TyrTS) from Bacillus stearothermophilus recognizes tRNA(Tyr), we constructed 14 new mutant TyrTS by site-directed mutagenesis, determined their kinetic properties and used these and previous data to construct a detailed structural model of the complex between TyrTS and the acceptor arm of tRNA(Tyr). In the model Arg207, Lys208, Asn 146 and Glu 152 interact with phosphate groups. A contact between guanine 1 and Trp 196 is unspecific. Adenine 73, the fourth base from the 3' end, is specifically recognized through Trp 196 and the main-chain carbonyl of Ala150. At the active site, adenine 76 might interact with Lys82 and Arg86. There is a tight complementarity in shape between the tRNA and the synthetase. TyrTS and tRNA(Tyr) form an additional contact, in the vicinity of adenine 73, when their complex goes from the initial state to the transition state. The rate of aminoacylation, through the precise recognition of adenine 73, could thus be an important factor of discrimination by TyrTS among tRNAs.
authors: Unité de Biochimie des Régulations Cellulaires (C N RS U RA D1129), Institut Pasteur, Paris, France.
authors: E
authors: E
authors: Labouze
authors: H
authors: H
authors: Bedouelle
mesh_terms: D000604:Amino Acyl-tRNA Synthetases / Q000235:genetics*; D001665:Binding Sites; D001411:Geobacillus stearothermophilus; D007700:Kinetics; D046911:Macromolecular Substances; D008958:Models, Molecular; D009154:Mutation; D012329:RNA, Bacterial / Q000235:genetics*; D012345:RNA, Transfer, Amino Acid-Specific / Q000235:genetics*; D012365:RNA, Transfer, Tyr / Q000235:genetics*; D014448:Tyrosine-tRNA Ligase / Q000235:genetics*
chemical_list: D046911:Macromolecular Substances; D012329:RNA, Bacterial; D012345:RNA, Transfer, Amino Acid-Specific; D012365:RNA, Transfer, Tyr; D000604:Amino Acyl-tRNA Synthetases; D014448:Tyrosine-tRNA Ligase