Kinetic properties of Escherichia coli ribosomes with altered forms of S12

J Mol Biol. 1992 Apr 20;224(4):1011-27. doi: 10.1016/0022-2836(92)90466-w.

Abstract

E. coli ribosomes with alterations in S12 leading to streptomycin resistance (SmR), dependence (SmD) and pseudodependence (SmP) were studied with the quench-flow technique. Kinetic changes at the various steps of the elongation cycle were identified. The rate of hydrolysis of GTP in the ternary complex in the ribosomal A-site is decreased drastically in SmD and moderately in SmP in relation to wild-type ribosomes. Addition of streptomycin restores much of the wild-type behaviour. The SmD, SmP and SmR ribosomes have an enhanced GTP-hydrolysis idling reaction on EF-Tu, which is correlated with how aggressive proofreaders these ribosomes are in steady-state assays. We use our in vitro findings to discuss the in vivo physiology of these mutants as well as mechanistic features of E. coli translation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Escherichia coli / metabolism
  • GTP Phosphohydrolase-Linked Elongation Factors / metabolism
  • Guanosine Triphosphate / metabolism
  • Kinetics
  • Mutation
  • Peptide Chain Elongation, Translational*
  • Peptide Elongation Factor Tu / metabolism
  • Peptidyl Transferases / metabolism
  • Ribosomal Proteins / metabolism*
  • Ribosomes / metabolism*
  • Ribosomes / ultrastructure
  • Streptomycin / pharmacology

Substances

  • Ribosomal Proteins
  • ribosomal protein S12
  • Guanosine Triphosphate
  • Peptidyl Transferases
  • GTP Phosphohydrolase-Linked Elongation Factors
  • Peptide Elongation Factor Tu
  • Streptomycin