Identification of quorum sensing-related regulons in Vibrio vulnificus by two-dimensional gel electrophoresis and differentially displayed reverse transcriptase PCR

FEMS Immunol Med Microbiol. 2007 Jun;50(1):94-103. doi: 10.1111/j.1574-695X.2007.00236.x.

Abstract

Vibrio vulnificus is thought to employ a quorum-sensing system to control the expression of a global gene. In this study, proteomes and transcriptomes of a lacZ null mutant, VvSR Delta Z, and a luxS-smcR double mutant, VvSR Delta ZSR, were compared with the parent strain, VvAR, by means of two-dimensional gel electrophoresis (2D-PAGE) and differentially displayed reverse transcriptase PCR (DDRT-PCR). 2D-PAGE analysis showed that 36 protein spots were differentially expressed, 14 of which have been identified by peptide-mass fingerprinting. The expression of eight cellular proteins was repressed by luxS and smcR mutation: Zn-dependent protease, 6-phosophofructokinase, periplasmic ABC-type Fe3(+) transport system, deoxyribose-phosphate aldolase, phosphomannomutase, orotidine-5'-phosphate decarboxylase, uridylate kinase, and an unidentified protein. These proteins are involved in virulence, adaptation to environmental stress, biosynthesis of LPS, and cell multiplication. Phage shock protein A, a chemotaxis signal transduction protein, and an uncharacterized low-complexity protein were activated in the cellular components of the luxS-smcR mutant. However, only three proteins, of unknown function, were identified in the extracellular components of the mutants. Analysis of transcriptomes with DDRT-PCR showed that two genes, phosphoribosylformylglycinamidine synthase and ATP-dependent protease HslVU protease were regulated at the transcriptional level by luxS and smcR gene mutation. The results from this study show conclusively that luxS/smcR quorum sensing endows a global change in gene expression to V. vulnificus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / biosynthesis
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Carbon-Sulfur Lyases / biosynthesis
  • Carbon-Sulfur Lyases / genetics
  • Carbon-Sulfur Lyases / metabolism
  • Cell Cycle Proteins / biosynthesis
  • Cell Cycle Proteins / genetics
  • Cell Cycle Proteins / metabolism
  • Electrophoresis, Gel, Two-Dimensional / methods
  • Gene Expression Profiling / methods
  • Gene Expression Regulation, Bacterial
  • Proteomics / methods
  • Quorum Sensing / genetics
  • Quorum Sensing / physiology*
  • Regulon / genetics
  • Regulon / physiology*
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Transcription, Genetic
  • Vibrio vulnificus / genetics
  • Vibrio vulnificus / growth & development
  • Vibrio vulnificus / metabolism
  • Vibrio vulnificus / physiology*

Substances

  • Bacterial Proteins
  • Cell Cycle Proteins
  • SMC protein, Bacteria
  • Carbon-Sulfur Lyases
  • LuxS protein, Bacteria