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A protein phosphorylation module patterns the Bacillus subtilis spore outer coat.

Carolina Freitas1, Jarnaja Plannic1,2, Rachele Isticato3

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Phosphorylation of Bacillus subtilis spore coat protein CotG by kinase CotH is crucial for forming the outer coat

Keywords:
CotBCotGCotHprotein kinaseprotein phosphorylationspore coat

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Area of Science:

  • Microbiology and Molecular Biology
  • Bacterial spore formation
  • Protein phosphorylation

Background:

  • The Bacillus subtilis spore coat comprises over 80 self-organizing proteins forming distinct layers: basal, inner, outer, and crust.
  • CotB is a major component of the outer spore coat, with its C-terminal region (SKRB) being polyphosphorylated by the Ser/Thr kinase CotH.

Purpose of the Study:

  • To investigate the role of the coat protein CotG in the phosphorylation of CotB and its impact on Bacillus subtilis spore coat assembly.
  • To elucidate the specific contribution of CotG-CotB interactions and CotH-mediated phosphorylation to the structural organization of the outer spore coat.

Main Methods:

  • In vivo and heterologous system studies to analyze protein interactions and phosphorylation events.
  • Analysis of Bacillus subtilis spore coat structure in wild-type, mutant (cotG deletion, inactive CotH), and modified strains (CotB SKRB deletion).

Main Results:

  • CotG interacts with CotB's C-terminus (SKRB) and promotes its phosphorylation by CotH.
  • CotG phosphorylation by CotH is enhanced without CotB; spores lacking functional CotH or CotG exhibit defects in outer coat striations but retain the crust.
  • Deletion of the CotB SKRB region does not significantly alter outer coat structure, highlighting the critical role of CotG phosphorylation.

Conclusions:

  • CotG-mediated promotion of CotB phosphorylation by CotH is essential for establishing the characteristic electron-dense striations of the Bacillus subtilis outer spore coat.
  • This protein phosphorylation module (CotB/CotH/CotG) is likely vital for spore surface layer morphogenesis in B. subtilis and related species.