Transcriptional regulation of cpsQ-mfpABC and mfpABC by CalR in Vibrio parahaemolyticus

Microbiologyopen. 2017 Aug;6(4):e00470. doi: 10.1002/mbo3.470. Epub 2017 Mar 20.

Abstract

The cpsQ-mfpABC locus is transcribed as two operons, i.e., cpsQ-mfpABC and mfpABC, in Vibrio parahaemolyticus, and both of them are all required for biofilm formation. CalR belongs to the LysR-type transcriptional regulator family, and was originally identified as a repressor of the swarming motility and T3SS1 genes expression in V. parahaemolyticus. In the present work, a combination of qRT-PCR, primer extension, LacZ fusion expression, electrophoretic mobility shift assay, and DNase I footprinting assays were employed to elucidate the regulatory mechanisms of cpsQ-mfpABC and mfpABC by CalR. One transcription start site for each operon was detected and their activities were activated by CalR. His-CalR protected two DNA regions upstream of mfpABC against DNase I digestion, but no binding sites were detected in the promoter region of cpsQ-mfpABC, suggesting a direct and an indirect regulatory manner for mfpABC and cpsQ-mfpABC transcription by CalR, respectively. Collectively, the results presented here confirmed a new physiological role for CalR that acts as an activator for cpsQ-mfpABC and mfpABC transcription.

Keywords: Vibrio parahaemolyticus; cpsQ-mfpABC; CalR; regulation.

MeSH terms

  • Bacterial Proteins / biosynthesis*
  • Bacterial Proteins / genetics
  • Gene Expression Regulation, Bacterial*
  • Operon
  • Transcription Factors / metabolism*
  • Transcription Initiation Site
  • Transcription, Genetic*
  • Vibrio parahaemolyticus / genetics*
  • Vibrio parahaemolyticus / metabolism

Substances

  • Bacterial Proteins
  • Transcription Factors