title: Recognition by tryptophanyl-tRNA synthetases of discriminator base on tRNATrp from three biological domains.
abstract: To study the recognition by tryptophanyl-tRNA synthetase (TrpRS) of tRNA(Trp) discriminator base, mutations were introduced into the discriminator base of Bacillus subtilis, Archeoglobus fulgidus, and bovine tRNA(Trp), representing the three biological domains. When B. subtilis, A. fulgidus, and human TrpRS were used to acylate these tRNA(Trp), two distinct preference profiles regarding the discriminator base of different tRNA(Trp) substrates were found: G>A>U>C for B. subtilis TrpRS, and A>C>U>G for A. fulgidus and human TrpRS. The preference for G73 in tRNA(Trp) by bacterial TrpRS is much stronger than the modest preferences for A73 by the archaeal and eukaryotic TrpRS. Cross-species reactivities between TrpRS and tRNA(Trp) from the three domains were in accordance with the view that the evolutionary position of archaea is intermediate between those of eukarya and bacteria. NMR spectroscopy revealed that mutation of A73 to G73 in bovine tRNA(Trp) elicited a conformational alteration in the G1-C72 base pair. Mutation of G1-C72 to A1-U72 or disruption of the G1-C72 base pair also caused reduction of Trp-tRNA(Trp) formation. These observations identify a tRNA(Trp) structural region near the end of acceptor stem comprising A73 and G1-C72 as a crucial domain required for effective recognition by human TrpRS.
authors.affiliation: Department of Biochemistry, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong SAR, China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Guo
authors.forename: Qingguo
authors.initials: Q
authors.lastname: Gong
authors.forename: Ka-Lok
authors.initials: KL
authors.lastname: Tong
authors.forename: Bente
authors.initials: B
authors.lastname: Vestergaard
authors.forename: Annie
authors.initials: A
authors.lastname: Costa
authors.forename: Jean
authors.initials: J
authors.lastname: Desgres
authors.forename: Mansim
authors.initials: M
authors.lastname: Wong
authors.forename: Henri
authors.initials: H
authors.lastname: Grosjean
authors.forename: Guang
authors.initials: G
authors.lastname: Zhu
authors.forename: J Tze-Fei
authors.initials: JT
authors.lastname: Wong
authors.forename: Hong
authors.initials: H
authors.lastname: Xue
mesh_terms: D000818:Animals; D001483:Base Sequence; D002417:Cattle; D003001:Cloning, Molecular; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009154:Mutation; D009690:Nucleic Acid Conformation; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D012364:RNA, Transfer, Trp / Q000235:genetics*; D013997:Time Factors; D014364:Tryptophan / Q000737:chemistry*; D014369:Tryptophan-tRNA Ligase / Q000378:metabolism*
chemical_list: D012364:RNA, Transfer, Trp; D014364:Tryptophan; D014369:Tryptophan-tRNA Ligase