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Related Experiment Videos

flaD (sinR) mutations affect SigD-dependent functions at multiple points in Bacillus subtilis

M H Rashid1, J Sekiguchi

  • 1Department of Applied Biology, Faculty of Textile Science and Technology, Shinshu University, Nagano, Japan.

Journal of Bacteriology
|November 1, 1996
PubMed
Summary
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A flaD (sinR) mutation impacts sigD-lacZ expression and cell morphology. Overproducing sigmaD partially or fully restored autolysin synthesis but not flagellin or motility in mutants.

Area of Science:

  • Microbiology
  • Bacterial Genetics
  • Cellular Biology

Background:

  • The sigmaD (σD) factor regulates motility and flagellar synthesis in bacteria.
  • The sinR gene, encoding a repressor of σD, is crucial for regulating σD activity.
  • Mutations in flaD, which affect sinR function, can alter σD-dependent gene expression.

Purpose of the Study:

  • To investigate the specific roles of flaD mutations in regulating sigD-lacZ expression and downstream phenotypes.
  • To determine the effect of overproducing sigmaD on the filamentous phenotype and autolysin synthesis in flaD mutants.
  • To assess the impact of flaD mutations on flagellin synthesis and bacterial motility.

Main Methods:

  • Construction and characterization of flaD (sinR) null and flaD1 point mutations.

Related Experiment Videos

  • Analysis of sigD-lacZ reporter gene expression in wild-type and mutant strains.
  • Introduction of a sigmaD-overproducing plasmid (pHYSigD) into mutant strains.
  • Assessment of cell morphology, autolysin synthesis, flagellin synthesis, and motility.

    • Main Results:

    • A flaD (sinR) null mutation caused a 2-4 fold decrease in sigD-lacZ expression, while a flaD1 point mutation nearly abolished it.
    • Overproduction of sigmaD using pHYSigD corrected the filamentous phenotype in both mutant strains.
    • Autolysin synthesis was partially restored in the flaD1 mutant and completely restored in the flaD (sinR) null mutant.
    • Flagellin synthesis and motility remained unaffected in both mutant strains, even with sigmaD overproduction.

    Conclusions:

    • The flaD1 mutation severely impairs sigD-lacZ expression, suggesting a critical role in sigmaD regulation.
    • SigmaD overproduction can rescue the morphological defects and restore autolysin synthesis in flaD mutants.
    • Despite restored autolysin, flagellin synthesis and motility are not recovered, indicating complex regulatory pathways beyond sigmaD levels.