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Recognition of bases in Escherichia coli tRNA(Gln) by glutaminyl-tRNA synthetase: a complete identity set.

1992; Springer Nature; Volume: 11; Issue: 11 Linguagem: Inglês

10.1002/j.1460-2075.1992.tb05509.x

1460-2075

Yoji Hayase, Martina Jahn, Michael J. Rogers, Lee A. Sylvers, M. Koizumi, H. Inoue, Eiko Ohtsuka, Dieter Söll,

Enzyme Structure and Function

Research Article1 November 1992free access Recognition of bases in Escherichia coli tRNA(Gln) by glutaminyl-tRNA synthetase: a complete identity set. Y. Hayase Y. Hayase Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06511. Search for more papers by this author M. Jahn M. Jahn Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06511. Search for more papers by this author M.J. Rogers M.J. Rogers Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06511. Search for more papers by this author L.A. Sylvers L.A. Sylvers Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06511. Search for more papers by this author M. Koizumi M. Koizumi Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06511. Search for more papers by this author H. Inoue H. Inoue Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06511. Search for more papers by this author E. Ohtsuka E. Ohtsuka Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06511. Search for more papers by this author D. Söll D. Söll Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06511. Search for more papers by this author Y. Hayase Y. Hayase Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06511. Search for more papers by this author M. Jahn M. Jahn Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06511. Search for more papers by this author M.J. Rogers M.J. Rogers Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06511. Search for more papers by this author L.A. Sylvers L.A. Sylvers Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06511. Search for more papers by this author M. Koizumi M. Koizumi Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06511. Search for more papers by this author H. Inoue H. Inoue Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06511. Search for more papers by this author E. Ohtsuka E. Ohtsuka Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06511. Search for more papers by this author D. Söll D. Söll Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06511. Search for more papers by this author Author Information Y. Hayase1, M. Jahn1, M.J. Rogers1, L.A. Sylvers1, M. Koizumi1, H. Inoue1, E. Ohtsuka1 and D. Söll1 1Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06511. The EMBO Journal (1992)11:4159-4165https://doi.org/10.1002/j.1460-2075.1992.tb05509.x PDFDownload PDF of article text and main figures. ToolsAdd to favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinked InMendeleyWechatReddit Figures & Info The fidelity of protein biosynthesis rests largely on the correct aminoacylation of transfer RNAs by their cognate aminoacyl-tRNA synthetases. Previous studies have demonstrated that the interaction of Escherichia coli tRNA(Gln) with glutaminyl-tRNA synthetase (GlnRS) provides an excellent system for studying the basis of this highly specific recognition process. Correct aminoacylation depends on the set of nucleotides (identity elements) in tRNA(Gln) responsible for correct interaction with GlnRS. Specific contacts between tRNA(Gln) and GlnRS include the 2-amino group of guanosines. Therefore, we made a set of tRNA(Gln) variants in which specific guanosines were replaced by inosine using recombinant RNA technology. This resulted in a set of tRNAs that varied by single deletions of the amino group from guanine residues, thus allowing us to test the functional importance of these contacts. In addition, a number of mutants were made by transcription of mutated tRNA genes with base changes at position 10, 16 or 25. In vitro aminoacylation of these mutants showed decreases in the specificity constant (kcat/KM) of up to 300-fold, with kcat being the parameter most affected. These experiments reveal G10 as a new element of glutamine identity. In addition, the interaction of G2, G3 and G10 with GlnRS via the 2-amino group is significant for tRNA discrimination. Based on these results, and on earlier data, we propose a complete set of bases as identity elements for tRNA(Gln). Previous ArticleNext Article Volume 11Issue 111 November 1992In this issue RelatedDetailsLoading ...