title: ZpdN, a Plasmid-Encoded Sigma Factor Homolog, Induces pBS32-Dependent Cell Death in Bacillus subtilis.
abstract: The ancestral Bacillus subtilis strain 3610 contains an 84-kb plasmid called pBS32 that was lost during domestication of commonly used laboratory derivatives. Here we demonstrate that pBS32, normally present at 1 or 2 copies per cell, increases in copy number nearly 100-fold when cells are treated with the DNA-damaging agent mitomycin C. Mitomycin C treatment also caused cell lysis dependent on pBS32-borne prophage genes. ZpdN, a sigma factor homolog encoded by pBS32, was required for the plasmid response to DNA damage, and artificial expression of ZpdN was sufficient to induce pBS32 hyperreplication and cell death. Plasmid DNA released by cell death was protected by the capsid protein ZpbH, suggesting that the plasmid was packaged into a phagelike particle. The putative particles were further indicated by CsCl sedimentation but were not observed by electron microscopy and were incapable of killing B. subtilis cells extracellularly. We hypothesize that pBS32-mediated cell death releases a phagelike particle that is defective and unstable.
  
  
  
  Prophages are phage genomes stably integrated into the host bacterium's chromosome and less frequently are maintained as extrachromosomal plasmids. Here we report that the extrachromosomal plasmid pBS32 of Bacillus subtilis encodes a prophage that, when activated, kills the host. pBS32 also encodes both the sigma factor homolog ZpdN that is necessary and sufficient for prophage induction and the protein ComI, which is a potent inhibitor of DNA uptake by natural transformation. We provide evidence that the entire pBS32 sequence may be part of the prophage and thus that competence inhibition may be linked to lysogeny.
authors.affiliation: Indiana University, Department of Biology, Bloomington, Indiana, USA.
authors.forename: B-E
authors.initials: BE
authors.lastname: Myagmarjav
authors.affiliation: Indiana University, Department of Biology, Bloomington, Indiana, USA.
authors.forename: M A
authors.initials: MA
authors.lastname: Konkol
authors.affiliation: Indiana University, Department of Biology, Bloomington, Indiana, USA.
authors.forename: J
authors.initials: J
authors.lastname: Ramsey
authors.affiliation: Indiana University, Department of Biology, Bloomington, Indiana, USA.
authors.forename: S
authors.initials: S
authors.lastname: Mukhopadhyay
authors.affiliation: Indiana University, Department of Biology, Bloomington, Indiana, USA dbkearns@indiana.edu.
authors.forename: D B
authors.initials: DB
authors.lastname: Kearns
mesh_terms: D001412:Bacillus subtilis / Q000166:cytology* / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D018628:Gene Dosage; D010957:Plasmids / Q000235:genetics* / Q000378:metabolism; D012808:Sigma Factor / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D012808:Sigma Factor