Functional analysis of SoxR residues affecting transduction of oxidative stress signals into gene expression

J Biol Chem. 2004 Oct 1;279(40):41603-10. doi: 10.1074/jbc.M405512200. Epub 2004 Jul 29.

Abstract

SoxR protein, a member of the MerR family of transcriptional activators, mediates a global oxidative stress response in Escherichia coli. Upon oxidation or nitrosylation of its [2Fe-2S] centers SoxR activates its target gene, soxS, by mediating a structural transition in the promoter DNA that stimulates initiation by RNA polymerase. We explored the molecular basis of this signal transduction by analyzing mutant SoxR proteins defective in responding to oxidative stress signals in vivo.We have confirmed that the DNA binding domain of SoxR is highly conserved compared with other MerR family proteins and functions in a similar manner to activate transcription. Several mutations in the dimerization domain of SoxR disrupted intersubunit communication, and the resulting proteins were unable to propagate redox signals to the soxS promoter. Mutations scattered throughout the polypeptide yielded proteins that were under-responsive to in vivo redox signals, which indicates that the redox properties of the [2Fe-2S] centers are influenced by global protein structure. These findings indicate that SoxR functions as a redox-responsive molecular switch in which subunit interactions transduce a subtle alteration in oxidation state into a profound change in DNA structure.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / physiology
  • Escherichia coli Proteins / physiology
  • Gene Expression Regulation, Bacterial*
  • Models, Molecular
  • Mutation
  • Oxidation-Reduction
  • Oxidative Stress*
  • Protein Conformation
  • Sequence Alignment
  • Signal Transduction*
  • Transcription Factors / chemistry*
  • Transcription Factors / physiology
  • Transcription, Genetic

Substances

  • Bacterial Proteins
  • Escherichia coli Proteins
  • Transcription Factors
  • SoxR protein, Bacteria