Exploring the aminoacylation function of transfer RNA by macromolecular engineering approaches. Involvement of conformational features in the charging process of yeast tRNA(Asp)

Biochimie
R GiegéJ P Ebel

Abstract

This report presents the conceptual and methodological framework that presently underlies the experiments designed to decipher the structural features in tRNA important for its aminoacylation by aminoacyl-tRNA synthetases. It emphasizes the importance of conformational features in tRNA for an optimized aminoacylation. This is illustrated by selected examples on yeast tRNA(Asp). Using the phage T7 transcriptional system, a series of tRNA(Asp) variants were created in which conformational elements were modified. It is shown that aspartyl-tRNA synthetase tolerates conformational variability in tRNA(Asp) at the level of the D-loop and variable region, of the tertiary Levitt base-pair 15-48 which can be inverted and in the T-arm in which residue 49 can be excised. However, changing the anticodon region completely abolishes the aspartylation capacity of the variants. Transplanting the phenylalanine identity elements into a different tRNA(Asp) variant presenting conformational characteristics of tRNA(Phe) converts this molecule into a phenylalanine acceptor but is less efficient than wild-type tRNA(Phe). This engineered tRNA completely loses its aspartylation capacity, showing that some aspartic acid and phenylalanine identity determi...Continue Reading

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Citations

Jan 1, 1996·Biochimie·R GiegéD Moras
Jun 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·J NormanlyJ Abelson
Apr 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·M J RogersD Söll
Jul 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·Y M HouR Giegé
Sep 1, 1996·Nucleic Acids Research·C E HolmesR Giegé
May 5, 1992·Biochemistry·Y M Hou, P Schimmel
Jun 12, 2019·IUBMB Life·Catherine Florentz, Richard Giegé
Jul 27, 1993·Biochemistry·G A Komatsoulis, J Abelson
Jan 14, 1994·Science·M E SaksJ N Abelson

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