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PMID: 1377381 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S. Corrected and Republished Article

Competition of aminoacyl-tRNA synthetases for tRNA ensures the accuracy of aminoacylation.

Nucleic acids research ·Vol. 20 ·No. 11 ·1992-00-11 ·Pages 2847-52

Sherman JM, Rogers MJ, Söll D

Abstract

The accuracy of protein biosynthesis rests on the high fidelity with which aminoacyl-tRNA synthetases discriminate between tRNAs. Correct aminoacylation depends not only on identity elements (nucleotides in certain positions) in tRNA (1), but also on competition between different synthetases for a given tRNA (2). Here we describe in vivo and in vitro experiments which demonstrate how variations in the levels of synthetases and tRNA affect the accuracy of aminoacylation. We show in vivo that concurrent overexpression of Escherichia coli tyrosyl-tRNA synthetase abolishes misacylation of supF tRNA(Tyr) with glutamine in vivo by overproduced glutaminyl-tRNA synthetase. In an in vitro competition assay, we have confirmed that the overproduction mischarging phenomenon observed in vivo is due to competition between the synthetases at the level of aminoacylation. Likewise, we have been able to examine the role competition plays in the identity of a non-suppressor tRNA of ambiguous identity, tRNA(Glu). Finally, with this assay, we show that the identity of a tRNA and the accuracy with which it is recognized depend on the relative affinities of the synthetases for the tRNA. The in vitro competition assay represents a general method of obtaining qualitative information on tRNA identity in a competitive environment (usually only found in vivo) during a defined step in protein biosynthesis, aminoacylation. In addition, we show that the discriminator base (position 73) and the first base of the anticodon are important for recognition by E. coli tyrosyl-tRNA synthetase.

MeSH Terms
Amino Acyl-tRNA Synthetases/metabolism Binding, Competitive Escherichia coli Genes, Suppressor Glutamate-tRNA Ligase/metabolism RNA, Bacterial/metabolism RNA, Transfer, Gln/metabolism RNA, Transfer, Glu/metabolism RNA, Transfer, Tyr/metabolism Substrate Specificity Transfer RNA Aminoacylation Tyrosine-tRNA Ligase/metabolism
Chemicals
RNA, Bacterial RNA, Transfer, Gln RNA, Transfer, Glu RNA, Transfer, Tyr Amino Acyl-tRNA Synthetases Tyrosine-tRNA Ligase Glutamate-tRNA Ligase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Sherman J M
Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06511.
Rogers M J
Söll D
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/20.11.2847
Published
1992-00-11
Pages
2847-52
Language
English
Region
England
NLM ID
0411011
PMCID
PMC336931
Subset
IM
Corrections
RepublishedFrom
PMID: 16617497
External Links
PubMed source DOI: 10.1093/nar/20.11.2847 PMC Full Text View journal details
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