Identification of CgeA as a glycoprotein that anchors polysaccharides to the spore surface in Bacillus subtilis

Yusei Nakaya, Miu Uchiike, Mayuko Hattori, Momoka Moriyama, Kimihiro Abe, Ella Kim, Patrick Eichenberger, Daisuke Imamura, Tsutomu Sato

Research output: Contribution to journalArticlepeer-review


The Bacillus subtilis spore is composed of a core, containing chromosomal DNA, surrounded by a cortex layer made of peptidoglycan, and a coat composed of concentric proteinaceous layers. A polysaccharide layer is added to the spore surface, and likely anchored to the crust, the coat outermost layer. However, the identity of the coat protein(s) to which the spore polysaccharides (SPS) are attached is uncertain. First, we showed that the crust proteins CotVWXYZ and CgeA were all contained in the peeled SPS layer obtained from a strain missing CotE, the outer coat morphogenetic protein, suggesting that the SPS is indeed bound to at least one of the spore surface proteins. Second, CgeA is known to be located at the most downstream position in the crust assembly pathway. An analysis of truncated variants of CgeA suggested that its N-terminal half is required for localization to the spore surface, while its C-terminal half is necessary for SPS addition. Third, an amino acid substitution strategy revealed that SPS was anchored at threonine 112 (T112), which constitutes a probable O-glycosylation site on CgeA. Our results indicated that CgeA is a glycoprotein required to initiate SPS assembly and serves as an anchor protein linking the crust and SPS layers.

Original languageEnglish (US)
Pages (from-to)384-396
Number of pages13
JournalMolecular Microbiology
Issue number3
StatePublished - Sep 2023


  • Bacillus subtilis
  • glycosylation
  • sporulation

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology


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