Transfer RNA identity change in anticodon variants of E. Coli tRNAPhe in vivo

Hyun Soo Kim, Ick Young Kim, Dieter Söll, Se Yong Lee

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

The anticodon sequence is a major recognition element for most aminoacyl-tRNA synthetases. We investigated the in vivo effects of changing the anticodon on the aminoacylation specificity in the example of E. coli tRNAPhe. Constructing different anticodon mutants of E. coli tRNAPhe by site-directed mutagenesis, we isolated 22 anticodon mutant tRNAPhe; the anticodons corresponded to 16 amino acids and an opal stop codon. To examine whether the mutant tRNAs had changed their amino acid acceptor specificity in vivo, we tested the viability of E. coli strains containing these tRNAPhe genes in a medium which permitted tRNA induction. Fourteen mutant tRNA genes did not affect host viability. However, eight mutant tRNA genes were toxic to the host and prevented growth, presumably because the anticodon mutants led to translational errors. Many mutant tRNAs which did not affect host viability were not aminoacylated in vivo. Three mutant tRNAs containing anticodon sequences corresponding to lysine (UUU), methionine (CAU) and threonine (UGU) were charged with the amino acid corresponding to their anticodon, but not with phenylalanine. These three tRNAs and tRNAPhe are located in the same cluster in a sequence similarity dendrogram of total E. coli tRNAs. The results support the idea that such tRNAs arising from in vivo evolution are derived by anticodon change from the same ancestor tRNA.

Original languageEnglish
Pages (from-to)76-82
Number of pages7
JournalMolecules and Cells
Volume10
Issue number1
Publication statusPublished - 2000 Feb 29

Fingerprint

RNA, Transfer, Phe
Anticodon
Transfer RNA
Escherichia coli
Amino Acids
Genes
Aminoacylation
Amino Acyl-tRNA Synthetases
Terminator Codon
Poisons
Threonine
Site-Directed Mutagenesis
Phenylalanine
Methionine
Lysine

Keywords

  • Anticodon
  • Evolution
  • Identity
  • tRNA

ASJC Scopus subject areas

  • Cell Biology
  • Genetics
  • Molecular Biology

Cite this

Transfer RNA identity change in anticodon variants of E. Coli tRNAPhe in vivo. / Kim, Hyun Soo; Kim, Ick Young; Söll, Dieter; Lee, Se Yong.

In: Molecules and Cells, Vol. 10, No. 1, 29.02.2000, p. 76-82.

Research output: Contribution to journalArticle

Kim, Hyun Soo ; Kim, Ick Young ; Söll, Dieter ; Lee, Se Yong. / Transfer RNA identity change in anticodon variants of E. Coli tRNAPhe in vivo. In: Molecules and Cells. 2000 ; Vol. 10, No. 1. pp. 76-82.
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