Single-base mutations at position 2661 of Escherichia coli 23S rRNA increase efficiency of translational proofreading

P. Melancon, W. E. Tapprich, L. Brakier-Gingras

Research output: Contribution to journalArticlepeer-review

29 Scopus citations

Abstract

Two single-base substitutions were constructed in the 2660 loop of Escherichia coli 23S rRNA (G2661→C or U) and were introduced into the rrnB operon cloned in plasmid pKK3535. Ribosomes were isolated from bacteria transformed with the mutated plasmids and assayed in vitro in a poly(U)- directed system for their response to the misreading effect of streptomycin, neomycin, and gentamicin, three aminoglycoside antibiotics known to impair the proofreading control of translational accuracy. Both mutations decreased the stimulation of misreading by these drugs, but neither interfered with their binding to the ribosome. The response of the mutant ribosomes to these drugs suggests that the 2660 loop, which belongs to the elongation factor Tu binding site, is involved in the proofreading step of the accuracy control. In vivo, both mutations reduced read-through of nonsense codons and frameshifting, which can also be related to the increased efficiency in proofreading control which they confer to ribosomes.

Original languageEnglish (US)
Pages (from-to)7896-7901
Number of pages6
JournalJournal of bacteriology
Volume174
Issue number24
DOIs
StatePublished - 1992
Externally publishedYes

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

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