Published online 27 June 2006
© 2006 The Author(s)
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A base pair at the bottom of the anticodon stem is reciprocally preferred for discrimination of cognate tRNAs by Escherichia coli lysyl- and glutaminyl-tRNA synthetases
Department of Biomolecular Science, Faculty of Engineering, Gifu University, 1-1 Yanagido Gifu 501-1193, Japan
*To whom correspondence should be addressed. Tel: +81 58 293 2644; Fax: +81 58 230 1893; Email: don{at}biomol.gifu-u.ac.jp
Received March 7, 2006. Revised May 23, 2006. Accepted May 25, 2006.
Although the yeast amber suppressor tRNATyr is a good candidate for a carrier of unnatural amino acids into proteins, slight misacylation with lysine was found to occur in an Escherichia coli protein synthesis system. Although it was possible to restrain the mislysylation by genetically engineering the anticodon stem region of the amber suppressor tRNATyr, the mutant tRNA showing the lowest acceptance of lysine was found to accept a trace level of glutamine instead. Moreover, the glutamine-acceptance of various tRNATyr transcripts substituted at the anticodon stem region varied in reverse proportion to the lysine-acceptance, similar to a ‘seesaw’. The introduction of a C31–G39 base pair at the site was most effective for decreasing the lysine-acceptance and increasing the glutamine-acceptance. When the same substitution was introduced into E.coli tRNALys transcripts, the lysine-accepting activity was decreased by 100-fold and faint acceptance of glutamine was observed. These results may support the idea that there are some structural element(s) in the anticodon stem of tRNA, which are not shared by aminoacyl-tRNA synthetases that have similar recognition sites in the anticodon, such as E.coli lysyl- and glutaminyl-tRNA synthetases.
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