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Related Experiment Videos

Two ResD-controlled promoters regulate ctaA expression in Bacillus subtilis.

S Paul1, X Zhang, F M Hulett

  • 1Laboratory for Molecular Biology, Department of Biological Sciences, University of Illinois at Chicago, 900 S. Ashland Ave., Chicago, IL 60607, USA.

Journal of Bacteriology
|April 28, 2001
PubMed
Summary

The Bacillus subtilis ResDE system regulates respiration genes. ResD~P activates the ctaA gene promoter, with distinct promoters (Pv and Ps) active at different growth phases, requiring specific ResD binding sites and sigma factors.

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Gene Regulation

Background:

  • The Bacillus subtilis ResDE two-component system is crucial for regulating genes involved in aerobic and anaerobic respiration.
  • The ctaA gene, essential for aerobic respiration, requires ResD for its expression in vivo.
  • The ctaAB promoter region contains three ResD binding sites (A1, A2, A3) influencing ctaA expression.

Purpose of the Study:

  • To investigate the role of ResD~P in activating the ctaA promoter using an in vitro transcription system.
  • To identify the specific ResD binding sites and sigma factors involved in ctaA transcription.
  • To determine the temporal regulation of ctaA gene expression during different growth phases.

Main Methods:

  • In vitro transcription assays using purified RNA polymerase (RNAP) and ResD~P.

Related Experiment Videos

  • Analysis of promoter activity with different ResD binding site deletions.
  • Characterization of RNAP isolated from various growth phases and mutant strains (spo0A, sigE).

    • Main Results:

    • The ctaA promoter exhibits two transcriptional start sites (Pv and Ps).
    • Binding site A2 is sufficient for weak transcription from Pv by Esigma(A), enhanced by ResD~P.
    • Maximal transcription from Ps requires binding sites A2 and A3, T(4) RNAP, and potentially Esigma(E).

    Conclusions:

    • ResD~P plays a key role in activating the ctaA promoter.
    • The ctaA gene is transcribed from Pv during late exponential to early stationary phases, requiring ResD binding site A2, Esigma(A), and ResD~P.
    • Transcription from Ps occurs during later stationary phases, dependent on binding sites A2 and A3, ResD~P, and Esigma(E) or a SigE-dependent sigma factor.