The ribosome-bound chaperones RAC and Ssb1/2p are required for accurate translation in Saccharomyces cerevisiae

Mol Cell Biol. 2004 Oct;24(20):9186-97. doi: 10.1128/MCB.24.20.9186-9197.2004.

Abstract

The chaperone homologs RAC (ribosome-associated complex) and Ssb1/2p are anchored to ribosomes; Ssb1/2p directly interacts with nascent polypeptides. The absence of RAC or Ssb1/2p results in a similar set of phenotypes, including hypersensitivity against the aminoglycoside paromomycin, which binds to the small ribosomal subunit and compromises the fidelity of translation. In order to understand this phenomenon we measured the frequency of translation termination and misincorporation in vivo and in vitro with a novel reporter system. Translational fidelity was impaired in the absence of functional RAC or Ssb1/2p, and the effect was further enhanced by paromomycin. The mutant strains suffered primarily from a defect in translation termination, while misincorporation was compromised to a lesser extent. Consistently, a low level of soluble translation termination factor Sup35p enhanced growth defects in the mutant strains. Based on the combined data we conclude that RAC and Ssb1/2p are crucial in maintaining translational fidelity beyond their postulated role as chaperones for nascent polypeptides.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphatases
  • Amino Acid Sequence
  • Anti-Bacterial Agents / pharmacology
  • Base Sequence
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • Gene Expression Regulation, Fungal
  • Genes, Reporter
  • HSP70 Heat-Shock Proteins / genetics
  • HSP70 Heat-Shock Proteins / metabolism*
  • Macromolecular Substances
  • Molecular Chaperones / genetics
  • Molecular Chaperones / metabolism*
  • Molecular Sequence Data
  • Paromomycin / pharmacology
  • Peptide Termination Factors / metabolism
  • Point Mutation
  • Protein Binding
  • Protein Biosynthesis* / drug effects
  • Ribosomes / metabolism*
  • Saccharomyces cerevisiae / drug effects
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism*

Substances

  • Anti-Bacterial Agents
  • DNA-Binding Proteins
  • HSP70 Heat-Shock Proteins
  • Macromolecular Substances
  • Molecular Chaperones
  • Peptide Termination Factors
  • SSB1 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Paromomycin
  • Adenosine Triphosphatases
  • SSB2 protein, S cerevisiae