title: Selection of suppressor methionyl-tRNA synthetases: mapping the tRNA anticodon binding site.
abstract: Accurate aminoacylation of a tRNA by Escherichia coli methionyl-tRNA synthetase (MTS) is specified by the CAU anticodon. A genetic screening procedure was designed to isolate MTS mutants able to aminoacylate a methionine amber tRNA (CUA anticodon). Selected suppressor MTS enzymes all possess one or several mutations in the vicinity of Trp-461, a residue that is the major contributor to the stability of complexes formed with tRNAs having the cognate CAU anticodon. Analysis of catalytic properties of purified suppressor enzymes shows that they have acquired an additional specificity toward the amber anticodon without complete disruption of the methionine anticodon site. It is concluded that both positive and negative discrimination toward the binding of tRNA anticodon sequences is restricted to a limited region of the synthetase, residues 451-467.
authors.affiliation: Laboratoire de Biochimie, Unité Associée 240 du Centre National de la Recherche Scientifique, Ecole Polytechnique, Palaiseau, France.
authors.forename: T
authors.initials: T
authors.lastname: Meinnel
authors.forename: Y
authors.initials: Y
authors.lastname: Mechulam
authors.forename: D
authors.initials: D
authors.lastname: Le Corre
authors.forename: M
authors.initials: M
authors.lastname: Panvert
authors.forename: S
authors.initials: S
authors.lastname: Blanquet
authors.forename: G
authors.initials: G
authors.lastname: Fayat
mesh_terms: D000926:Anticodon / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005798:Genes, Bacterial; D014644:Genetic Variation; D007700:Kinetics; D008718:Methionine-tRNA Ligase / Q000235:genetics* / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D012343:RNA, Transfer / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D013489:Suppression, Genetic
chemical_list: D000926:Anticodon; D012343:RNA, Transfer; D008718:Methionine-tRNA Ligase