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Related Experiment Videos

Sequence and characterization of Bacillus subtilis CheW.

D W Hanlon1, L M Márquez-Magaña, P B Carpenter

  • 1Department of Biochemistry, College of Medicine, University of Illinois, Urbana 61801.

The Journal of Biological Chemistry
|June 15, 1992
PubMed
Summary

Researchers identified a Bacillus subtilis gene homologous to E. coli CheW. Disrupting this gene impaired bacterial chemotaxis, affecting motility and response to chemical signals.

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Area of Science:

  • Microbiology
  • Bacterial Motility
  • Chemotaxis

Background:

  • CheW proteins are essential components of the bacterial chemotaxis signaling pathway.
  • Understanding CheW function in Bacillus subtilis provides insights into conserved microbial signaling mechanisms.

Purpose of the Study:

  • To characterize a novel Bacillus subtilis gene homologous to Escherichia coli CheW.
  • To investigate the role of this CheW homolog in bacterial chemotaxis and motility.

Main Methods:

  • Gene cloning, sequencing, and subcloning of the Bacillus subtilis open reading frame (ORF).
  • Verification of ORF expression using a bacteriophage T7 expression system in E. coli.
  • Gene inactivation via insertion of a chloramphenicol acetyltransferase cassette.

Related Experiment Videos

  • Phenotypic analysis of the mutant strain on motility and swarm plates.
  • Complementation studies using a plasmid expressing wild-type CheW.
  • Chemotaxis assays, including capillary assays, in response to chemoeffectors.

    • Main Results:

    • A 156-amino acid polypeptide was identified with 28.6% amino acid identity to E. coli CheW.
    • The Bacillus subtilis CheW homolog is essential for proper chemotaxis.
    • Mutants exhibited a smooth swimming bias in the absence of chemoeffectors and were defective on swarm plates.
    • Chemotaxis was significantly impaired in the mutant strain, showing only transient responses to attractants and repellents.

    Conclusions:

    • The characterized Bacillus subtilis ORF encodes a functional CheW homolog crucial for chemotaxis.
    • This finding highlights the conserved role of CheW in bacterial signal transduction pathways across different species.
    • The study provides a foundation for further investigation into the specific mechanisms of chemotaxis regulation in Bacillus subtilis.