title: Macromolecular recognition through electrostatic repulsion.
abstract: In the process of genetic translation, each aminoacyl-tRNA synthetase specifically aminoacylates its cognate tRNAs and rejects the 19 other species of tRNAs. A decrease in the specificity of this reaction can result in misincorporations of amino acids into proteins and be deleterious to the cell. In the case of tyrosyl-tRNA synthetase from Bacillus stearothermophilus, the change of residue Glu152 into Ala results in erroneous interactions with non-cognate tRNAs. To analyse how Glu152 contributes to the discrimination between tRNAs by tyrosyl-tRNA synthetase, 11 changes to this residue were created by mutagenesis. The misaminoacylations of tRNA(Phe) and tRNA(Val) with tyrosine in vitro (on a scale going from 1 to 30) and the toxicity of tyrosyl-tRNA synthetase in vivo (on a scale from 1 to 10(7)) increased in a correlated way when the nature of the side chain in position 152 varied from negatively charged to uncharged then to positively charged. The aminoacylation of tRNA(Tyr) was unaffected by the mutations. The results show that the role of Glu152 in the discrimination between tRNAs is purely negative, that it acts by electrostatic repulsion of non-cognate tRNAs and that this mechanism has been conserved throughout evolution.
authors.affiliation: Groupe d'Ingénierie des Protéines (CNRS URA 1129), Institut Pasteur, Paris, France.
authors.forename: H
authors.initials: H
authors.lastname: Bedouelle
authors.forename: R
authors.initials: R
authors.lastname: Nageotte
mesh_terms: D005075:Biological Evolution; D001411:Geobacillus stearothermophilus / Q000187:drug effects / Q000201:enzymology; D018698:Glutamic Acid / Q000378:metabolism; D046911:Macromolecular Substances; D017354:Point Mutation / Q000502:physiology; D012360:RNA, Transfer, Phe / Q000378:metabolism*; D012365:RNA, Transfer, Tyr / Q000378:metabolism; D012366:RNA, Transfer, Val / Q000378:metabolism*; D013379:Substrate Specificity; D046249:Transfer RNA Aminoacylation / Q000235:genetics*; D014443:Tyrosine / Q000378:metabolism; D014448:Tyrosine-tRNA Ligase / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D046911:Macromolecular Substances; D012360:RNA, Transfer, Phe; D012365:RNA, Transfer, Tyr; D012366:RNA, Transfer, Val; D018698:Glutamic Acid; D014443:Tyrosine; D014448:Tyrosine-tRNA Ligase