Quorum-Sensing Regulator OpaR Directly Represses Seven Protease Genes in Vibrio parahaemolyticus

Frontiers in Microbiology
San-Chi Chang, Chia-Yin Lee

Abstract

Proteases play a key role in numerous bacterial physiological events. Microbial proteases are used in the pharmaceutical industry and in biomedical applications. The genus Vibrio comprises protease-producing bacteria. Proteases transform polypeptides into shorter chains for easier utilization. They also function as a virulence factor in pathogens. The mechanism by which protease genes are regulated in Vibrio parahaemolyticus, an emerging world-wide human pathogen, however, still remains unclear. Quorum sensing is the communication system of bacteria. OpaR is the master quorum-sensing regulator in V. parahaemolyticus. In the present study, quantitative reverse transcriptase-polymerase chain reaction and protease gene promoter-fusion reporter assays revealed that OpaR represses seven protease genes-three metalloprotease genes and four serine protease genes-which are involved in environmental survival and bacterial virulence. Furthermore, the electrophoresis mobility shift assay demonstrated that OpaR is bound directly to the promoter region of each of the seven protease genes. DNase I footprinting identified the sequence of these OpaR-binding sites. ChIP-seq analyses revealed 435 and 835 OpaR-binding sites in the late-log and sta...Continue Reading

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Datasets Mentioned

BETA
GSE122479

Methods Mentioned

BETA
ChIP-seq
electrophoretic mobility shift assay
footprinting
electrophoresis
RNA-seq
ChIPseq
PCR
ChIP
Assay

Software Mentioned

SignalP
CLC Genomics Workbench
MEME
TOMTOM
ARNold
Multiple Expectation Maximization for Motif Elicitation ( MEME )
Peak Scanner

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