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Science 31 March 1995:
Vol. 267. no. 5206, pp. 1994 - 1996
DOI: 10.1126/science.7701322
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Articles

Science, Vol 267, Issue 5206, 1994-1996
Copyright © 1995 by American Association for the Advancement of Science

articles

Switching recognition of two tRNA synthetases with an amino acid swap in a designed peptide

DS Auld and P Schimmel

Department of Biology, Massachusetts Institute of Technology, Cambridge 02139, USA.

The genetic code is based on specific interactions between transfer RNA (tRNA) synthetases and their cognate tRNAs. The anticodons for methionine and isoleucine tRNAs differ by a single nucleotide, and changing this nucleotide in an isoleucine tRNA is sufficient to change aminoacylation specificity to methionine. Results of combinatorial mutagenesis of an anticodon-binding-helix loop peptide were used to design a hybrid sequence composed of amino acid residues from methionyl- and isoleucyl-tRNA synthetases. When the hybrid sequence was transplanted into isoleucyl-tRNA synthetase, active enzyme was generated in vivo and in vitro. The transplanted peptide did not confer function to methionyl-tRNA synthetase, but the substitution of a single amino acid within the transplanted peptide conferred methionylation and prevented isoleucylation. Thus, the swap of a single amino acid in the transplanted peptide switches specificity between anticodons that differ by one nucleotide.


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