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Genes encoding spore coat polypeptides from Bacillus subtilis.

W Donovan1, L B Zheng, K Sandman

  • 1Department of Cellular and Developmental Biology, Harvard University, Cambridge, MA 02138.

Journal of Molecular Biology
|July 5, 1987
PubMed
Summary
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Researchers identified four Bacillus subtilis spore coat protein genes (cotA, cotB, cotC, cotD). Mutants showed altered spore germination and pigment production, revealing gene functions in spore structure and development.

Area of Science:

  • Microbiology
  • Bacterial genetics
  • Spore formation

Background:

  • Bacillus subtilis endospores possess a complex proteinaceous coat essential for protection.
  • The precise genetic basis and functional roles of individual spore coat proteins remain largely uncharacterized.

Purpose of the Study:

  • To clone and characterize structural genes encoding Bacillus subtilis spore coat proteins.
  • To determine the chromosomal locations and genetic functions of these spore coat protein genes.

Main Methods:

  • Cloning of spore coat protein genes (cotA, cotB, cotC, cotD) using synthetic oligonucleotide probes.
  • Insertion of a chloramphenicol-resistance (cat) gene to create tagged, insertionally inactivated alleles.
  • Genetic mapping of the inactivated genes on the B. subtilis chromosome.

Related Experiment Videos

  • Phenotypic analysis of mutant spores, including germination rates and pigment production.

    • Main Results:

    • Four structural genes (cotA, cotB, cotC, cotD) encoding spore coat proteins were successfully cloned.
    • Genetic mapping placed these genes at distinct chromosomal locations (52°, 290°, 168°, 200°).
    • Mutants exhibited subtle phenotypes: cotD mutants showed slightly delayed germination, and cotA mutants lacked characteristic brown pigment, with cotA identified as the previously known 'pig' gene.

    Conclusions:

    • The identified cot genes encode essential spore coat proteins in Bacillus subtilis.
    • These genes play roles in spore coat integrity, germination efficiency, and pigmentation.
    • The study provides foundational genetic data for understanding Bacillus subtilis spore coat assembly and function.