Protein Aggregation Caused by Aminoglycoside Action Is Prevented by a Hydrogen Peroxide Scavenger

Mol Cell. 2012 Dec 14;48(5):713-22. doi: 10.1016/j.molcel.2012.10.001. Epub 2012 Oct 30.

Abstract

Protein mistranslation causes growth arrest in bacteria, mitochondrial dysfunction in yeast, and neurodegeneration in mammals. It remains poorly understood how mistranslated proteins cause such cellular defects. Here we demonstrate that streptomycin, a bactericidal aminoglycoside that increases ribosomal mistranslation, induces transient protein aggregation in wild-type Escherichia coli. We further determined the aggregated proteome using label-free quantitative mass spectrometry. To identify genes that reduce cellular mistranslation toxicity, we selected from an overexpression library protein products that increased resistance against streptomycin and kanamycin. The selected proteins were significantly enriched in members of the oxidation-reduction pathway. Overexpressing one of these proteins, alkyl hydroperoxide reductase subunit F (a protein defending bacteria against hydrogen peroxide), but not its inactive mutant suppressed aggregated protein formation upon streptomycin treatment and increased aminoglycoside resistance. This work provides in-depth analyses of an aggregated proteome caused by streptomycin and suggests that cellular defense against hydrogen peroxide lowers the toxicity of mistranslation.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aminoglycosides / pharmacology*
  • Anti-Bacterial Agents / pharmacology*
  • Drug Resistance, Multiple, Bacterial / genetics
  • Escherichia coli / drug effects*
  • Escherichia coli / genetics
  • Escherichia coli / growth & development
  • Escherichia coli / metabolism
  • Escherichia coli Proteins / biosynthesis*
  • Escherichia coli Proteins / chemistry
  • Escherichia coli Proteins / genetics
  • Free Radical Scavengers / pharmacology*
  • Gene Expression Regulation, Bacterial
  • Hydrogen Peroxide / metabolism*
  • Kanamycin / pharmacology
  • Mass Spectrometry
  • Microbial Viability / drug effects
  • Mutation
  • Oxidative Stress / drug effects*
  • Oxidative Stress / genetics
  • Peroxiredoxins / biosynthesis
  • Peroxiredoxins / genetics
  • Protein Biosynthesis / drug effects*
  • Protein Folding
  • Proteomics / methods
  • Ribosomes / drug effects
  • Ribosomes / metabolism
  • Streptomycin / pharmacology
  • Time Factors

Substances

  • Aminoglycosides
  • Anti-Bacterial Agents
  • Escherichia coli Proteins
  • Free Radical Scavengers
  • Kanamycin
  • Hydrogen Peroxide
  • Peroxiredoxins
  • ahpF protein, E coli
  • Streptomycin