Covariation of a specificity-determining structural motif in an aminoacyl-tRNA synthetase and a tRNA identity element

Biochemistry
S A Hawko, C S Francklyn

Abstract

Transfer RNA (tRNA) identity determinants help preserve the specificity of aminoacylation in vivo, and prevent cross-species interactions. Here, we investigate covariation between the discriminator base (N73) element in histidine tRNAs and residues in the histidyl-tRNA synthetase (HisRS) motif 2 loop. A model of the Escherichia coli HisRS--tRNA(His) complex predicts an interaction between the prokaryotic conserved glutamine 118 of the motif 2 loop and cytosine 73. The substitution of Gln 118 in motif 2 with glutamate decreased discrimination between cytosine and uracil some 50-fold, but left overall rates of adenylation and aminoacylation unaffected. By contrast, substitutions at neighboring Glu 115 and Arg 121 affected both adenylation and aminoacylation, consistent with their predicted involvement in both half-reactions. Additional evidence for the involvement of the motif 2 loop was provided by functional analysis of a hybrid Saccharomyces cerevisiae-- E. coli HisRS possessing the 11 amino acid motif 2 loop of the yeast enzyme. Despite an overall decreased activity of nearly 1000-fold relative to the E. coli enzyme, the chimera nevertheless exhibited a modest preference for the yeast tRNA(His) over the E. coli tRNA, and pref...Continue Reading

Citations

Sep 9, 2008·Molecular Biology and Evolution·Ignacio LuqueJesús A G Ochoa de Alda
Jan 15, 2002·Nucleic Acids Research·Jerome C ShielsAnne M Baranger
Feb 14, 2006·Nucleic Acids Research·David H Ardell, Siv G E Andersson
Dec 19, 2006·Journal of Bacteriology·Chunxia WangKelly P Williams
Sep 11, 2012·The Journal of Physical Chemistry. B·Mantu Santra, Biman Bagchi
Oct 31, 2016·Methods : a Companion to Methods in Enzymology·Jamie A AbbottChristopher S Francklyn
Jul 30, 2005·The Journal of Biological Chemistry·Karen S ChampagneChristopher S Francklyn
Mar 23, 2017·Cellular and Molecular Life Sciences : CMLS·Yi-Hsueh LeeChien-Chia Wang

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