Functional analysis of the ComK protein of Bacillus coagulans

PLoS One. 2013;8(1):e53471. doi: 10.1371/journal.pone.0053471. Epub 2013 Jan 3.

Abstract

The genes for DNA uptake and recombination in Bacilli are commonly regulated by the transcriptional factor ComK. We have identified a ComK homologue in Bacillus coagulans, an industrial relevant organism that is recalcitrant for transformation. Introduction of B. coagulans comK gene under its own promoter region into Bacillus subtilis comK strain results in low transcriptional induction of the late competence gene comGA, but lacking bistable expression. The promoter regions of B. coagulans comK and the comGA genes are recognized in B. subtilis and expression from these promoters is activated by B. subtilis ComK. Purified ComK protein of B. coagulans showed DNA-binding ability in gel retardation assays with B. subtilis- and B. coagulans-derived probes. These experiments suggest that the function of B. coagulans ComK is similar to that of ComK of B. subtilis. When its own comK is overexpressed in B. coagulans the comGA gene expression increases 40-fold, while the expression of another late competence gene, comC is not elevated and no reproducible DNA-uptake could be observed under these conditions. Our results demonstrate that B. coagulans ComK can recognize several B.subtilis comK-responsive elements, and vice versa, but indicate that the activation of the transcription of complete sets of genes coding for a putative DNA uptake apparatus in B. coagulans might differ from that of B. subtilis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Bacillus / metabolism*
  • Bacterial Proteins / genetics*
  • Cloning, Molecular
  • DNA, Bacterial / genetics
  • Gene Expression Regulation, Bacterial*
  • Genetic Engineering / methods
  • Genome, Bacterial
  • Genomics
  • Molecular Sequence Data
  • Nucleotides / genetics
  • Plasmids / metabolism
  • Promoter Regions, Genetic
  • Response Elements
  • Species Specificity
  • Transcription Factors / genetics*

Substances

  • Bacterial Proteins
  • DNA, Bacterial
  • Nucleotides
  • Transcription Factors
  • comK protein, Bacillus subtilis

Associated data

  • GENBANK/JX518619
  • GENBANK/JX518620
  • GENBANK/JX518621
  • GENBANK/JX518622
  • GENBANK/JX518623
  • GENBANK/JX518624

Grants and funding

Part of this work was financially supported by a SenterNovem subsidy IS044081. This project was carried out within the research programme of the Kluyver Centre for Genomics of Industrial Fermentation which is part of the Netherlands Genomics Initiative/Netherlands Organization for Scientific Research. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.