Contributions of crust proteins to spore surface properties in Bacillus subtilis

Bentley Shuster, Mark Khemmani, Kimihiro Abe, Xiaoyu Huang, Yusei Nakaya, Nina Maryn, Sally Buttar, Adriana N. Gonzalez, Adam Driks, Tsutomu Sato, Patrick Eichenberger

Research output: Contribution to journalArticlepeer-review


Surface properties, such as adhesion and hydrophobicity, constrain dispersal of bacterial spores in the environment. In Bacillus subtilis, these properties are influenced by the outermost layer of the spore, the crust. Previous work has shown that two clusters, cotVWXYZ and cgeAB, encode the protein components of the crust. Here, we characterize the respective roles of these genes in surface properties using Bacterial Adherence to Hydrocarbons assays, negative staining of polysaccharides by India ink and Transmission Electron Microscopy. We showed that inactivation of crust genes caused increases in spore relative hydrophobicity, disrupted the spore polysaccharide layer, and impaired crust structure and attachment to the rest of the coat. We also found that cotO, previously identified for its role in outer coat formation, is necessary for proper encasement of the spore by the crust. In parallel, we conducted fluorescence microscopy experiments to determine the full network of genetic dependencies for subcellular localization of crust proteins. We determined that CotZ is required for the localization of most crust proteins, while CgeA is at the bottom of the genetic interaction hierarchy.

Original languageEnglish (US)
Pages (from-to)825-843
Number of pages19
JournalMolecular Microbiology
Issue number3
StatePublished - Mar 2019

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology


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