SinR controls enterotoxin expression in Bacillus thuringiensis biofilms

PLoS One. 2014 Jan 31;9(1):e87532. doi: 10.1371/journal.pone.0087532. eCollection 2014.

Abstract

The entomopathogen Bacillus thuringiensis produces dense biofilms under various conditions. Here, we report that the transition phase regulators Spo0A, AbrB and SinR control biofilm formation and swimming motility in B. thuringiensis, just as they control biofilm formation and swarming motility in the closely related saprophyte species B. subtilis. However, microarray analysis indicated that in B. thuringiensis, in contrast to B. subtilis, SinR does not control an eps operon involved in exopolysaccharides production, but regulates genes involved in the biosynthesis of the lipopeptide kurstakin. This lipopeptide is required for biofilm formation and was previously shown to be important for survival in the host cadaver (necrotrophism). Microarray analysis also revealed that the SinR regulon contains genes coding for the Hbl enterotoxin. Transcriptional fusion assays, Western blots and hemolysis assays confirmed that SinR controls Hbl expression, together with PlcR, the main virulence regulator in B. thuringiensis. We show that Hbl is expressed in a sustained way in a small subpopulation of the biofilm, whereas almost all the planktonic population transiently expresses Hbl. The gene coding for SinI, an antagonist of SinR, is expressed in the same biofilm subpopulation as hbl, suggesting that hbl transcription heterogeneity is SinI-dependent. B. thuringiensis and B. cereus are enteric bacteria which possibly form biofilms lining the host intestinal epithelium. Toxins produced in biofilms could therefore be delivered directly to the target tissue.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacillus cereus / physiology
  • Bacillus thuringiensis / physiology*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Biofilms*
  • Enterotoxins / biosynthesis*
  • Enterotoxins / genetics
  • Gene Expression Regulation, Bacterial / physiology*
  • Regulon / physiology*

Substances

  • Bacterial Proteins
  • Enterotoxins
  • FlaD protein, Bacteria

Grants and funding

TD was funded by the ‘Direction Générale à L'Armement’, France and IB was funded by the ‘Région Ile de France’, France. AF, OAØ and ABK were funded by a project grant from the Norwegian Research Council through the FUGE II Programme (Channel 3 grant ; project nr. 183421). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.