Streptomycin interferes with conformational coupling between codon recognition and GTPase activation on the ribosome

Nat Struct Mol Biol. 2004 Apr;11(4):316-22. doi: 10.1038/nsmb742. Epub 2004 Mar 7.

Abstract

Aminoacyl-tRNAs (aa-tRNAs) are selected by the ribosome through a kinetically controlled induced fit mechanism. Cognate codon recognition induces a conformational change in the decoding center and a domain closure of the 30S subunit. We studied how these global structural rearrangements are related to tRNA discrimination by using streptomycin to restrict the conformational flexibility of the 30S subunit. The antibiotic stabilized aa-tRNA on the ribosome both with a cognate and with a near-cognate codon in the A site. Streptomycin altered the rates of GTP hydrolysis by elongation factor Tu (EF-Tu) on cognate and near-cognate codons, resulting in almost identical rates of GTP hydrolysis and virtually complete loss of selectivity. These results indicate that movements within the 30S subunit at the streptomycin-binding site are essential for the coupling between base pair recognition and GTP hydrolysis, thus modulating the fidelity of aa-tRNA selection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding Sites
  • Codon / chemistry
  • Codon / genetics*
  • Codon / metabolism*
  • Escherichia coli / drug effects
  • Escherichia coli / genetics
  • GTP Phosphohydrolases / chemistry
  • GTP Phosphohydrolases / genetics
  • GTP Phosphohydrolases / metabolism*
  • Kinetics
  • RNA, Transfer, Met / metabolism
  • Ribosomes / chemistry
  • Ribosomes / metabolism*
  • Streptomycin / pharmacology

Substances

  • Codon
  • RNA, Transfer, Met
  • fMet-tRNA(fMet)
  • GTP Phosphohydrolases
  • Streptomycin