The Role of α-CTD in the Genome-Wide Transcriptional Regulation of the Bacillus subtilis Cells

PLoS One. 2015 Jul 8;10(7):e0131588. doi: 10.1371/journal.pone.0131588. eCollection 2015.

Abstract

The amino acid sequence of the RNA polymerase (RNAP) α-subunit is well conserved throughout the Eubacteria. Its C-terminal domain (α-CTD) is important for the transcriptional regulation of specific promoters in both Escherichia coli and Bacillus subtilis, through interactions with transcription factors and/or a DNA element called the "UP element". However, there is only limited information regarding the α-CTD regulated genes in B. subtilis and the importance of this subunit in the transcriptional regulation of B. subtilis. Here, we established strains and the growth conditions in which the α-subunit of RNAP was replaced with a C-terminally truncated version. Transcriptomic and ChAP-chip analyses revealed that α-CTD deficiency reduced the transcription and RNAP binding of genes related to the utilization of secondary carbon sources, transition state responses, and ribosome synthesis. In E. coli, it is known that α-CTD also contributes to the expression of genes related to the utilization of secondary carbon sources and ribosome synthesis. Our results suggest that the biological importance of α-CTD is conserved in B. subtilis and E. coli, but that its specific roles have diversified between these two bacteria.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacillus subtilis / drug effects
  • Bacillus subtilis / genetics*
  • Bacillus subtilis / growth & development
  • Carbon / pharmacology
  • Chromatin Immunoprecipitation
  • DNA, Bacterial / metabolism
  • DNA-Directed RNA Polymerases / chemistry
  • DNA-Directed RNA Polymerases / genetics
  • DNA-Directed RNA Polymerases / metabolism*
  • Down-Regulation / drug effects
  • Down-Regulation / genetics
  • Gene Expression Regulation, Bacterial / drug effects
  • Genes, Bacterial
  • Genetic Loci
  • Genome, Bacterial*
  • Isopropyl Thiogalactoside / metabolism
  • Multiprotein Complexes / metabolism
  • Promoter Regions, Genetic
  • Protein Binding / drug effects
  • Ribosomes / drug effects
  • Ribosomes / metabolism
  • Sigma Factor / metabolism
  • Transcription Elongation, Genetic / drug effects
  • Transcription, Genetic* / drug effects
  • Transcriptional Activation / drug effects
  • Transcriptional Activation / genetics
  • Transcriptome / genetics
  • Up-Regulation / drug effects
  • Up-Regulation / genetics
  • Xylose / metabolism

Substances

  • DNA, Bacterial
  • Multiprotein Complexes
  • Sigma Factor
  • Isopropyl Thiogalactoside
  • Carbon
  • Xylose
  • DNA-Directed RNA Polymerases
  • RNA polymerase alpha subunit

Grants and funding

This work was supported by JSPS KAKENHI, Grant Numbers 23241062 (to TO, SI and NO) and JST Advanced Low Carbon Technology Research and Development Program, ALCA, (to NO). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.