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The RssB response regulator directly targets sigma(S) for degradation by ClpXP.

Y Zhou1, S Gottesman, J R Hoskins

  • 1Laboratory of Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892-4255, USA.

Genes & Development
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Regulated degradation of the sigma(S) subunit in E. coli is crucial for gene expression control. The protein RssB acts as a catalyst, delivering sigma(S) to the ClpXP protease for degradation.

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Area of Science:

  • Molecular Biology
  • Protein Degradation
  • Gene Regulation

Background:

  • The sigma(S) subunit of E. coli RNA polymerase is key for stationary phase and stress response gene expression.
  • Regulated degradation of sigma(S) is a critical control mechanism for its activity.
  • In vivo, sigma(S) degradation involves the ClpXP protease and the response regulator-like protein RssB.

Purpose of the Study:

  • To elucidate the in vitro mechanism of sigma(S) degradation by ClpXP.
  • To define the specific role of RssB in targeting sigma(S) for proteolysis.
  • To investigate the regulatory factors influencing sigma(S) turnover.

Main Methods:

  • Reconstruction of sigma(S) degradation using purified components in vitro.
  • Analysis of protein-protein interactions between sigma(S), RssB, and ClpXP.
  • Investigation of the role of acetyl phosphate and ATP in the degradation process.

Main Results:

  • RssB directly delivers sigma(S) to ClpXP and significantly stimulates its degradation.
  • Acetyl phosphate is required for RssB-mediated sigma(S) degradation, indicating RssB phosphorylation is essential.
  • RssB acts catalytically, participating in multiple rounds of sigma(S) degradation and showing substrate specificity.
  • Formation of a ternary complex (sigma(S)-RssB-ClpX) and a larger complex with ClpP is crucial for degradation.

Conclusions:

  • RssB is a specific targeting protein that delivers sigma(S) to ClpXP for regulated degradation.
  • This mechanism highlights a novel pathway for controlled protein turnover, regulated by signaling events.
  • The findings provide insights into the precise control of gene expression through protein degradation in bacteria.