A single base mutation at position 2661 in E.coli 23S ribosomal RNA affects the binding of ternary complex to the ribosome

W. E. Tapprich, A. E. Dahlberg

Research output: Contribution to journalArticlepeer-review

57 Scopus citations

Abstract

A single base substitution mutation from guanine to cytosine was constructed at position 2661 of Escherichia coli 23S rRNA and cloned into the rrnB operon of the multi-copy plasmid pKK3535. The mutant plasmid was transformed into E.coli to determine the effect of the mutation on cell growth as well as the structural and functional properties of the mutant ribosomes in vivo and in vitro. The results show that the mutant ribosomes have a slower elongation rate and an altered affinity for EF-Tu-tRNA-GTP ternary complex. This supports previous findings which indicated that position 2661 is part of a region of 23S rRNA that forms a recognition site for binding of the ternary complex in the ribosomal A site. Combinations of the 2661 mutation with various mutations in ribosomal protein S12 also demonstrate that elements of both ribosomal subunits work in concert to form this binding site.

Original languageEnglish (US)
Pages (from-to)2649-2655
Number of pages7
JournalEMBO Journal
Volume9
Issue number8
StatePublished - 1990
Externally publishedYes

Keywords

  • EF-Tu
  • Mutagenesis
  • Translation
  • tRNA

ASJC Scopus subject areas

  • General Biochemistry, Genetics and Molecular Biology
  • General Immunology and Microbiology
  • Molecular Biology
  • General Neuroscience

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