title: Identification of quorum sensing-related regulons in Vibrio vulnificus by two-dimensional gel electrophoresis and differentially displayed reverse transcriptase PCR.
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.
authors.affiliation: Department of Infectious Diseases, BK21 for Veterinary Science and KRF Zoonotic Priority Research Institute, College of Veterinary Medicine, Seoul National University, Korea.
authors.forename: Na-Ri
authors.initials: NR
authors.lastname: Shin
authors.forename: Deog Yong
authors.initials: DY
authors.lastname: Lee
authors.forename: Han Sang
authors.initials: HS
authors.lastname: Yoo
mesh_terms: D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D013437:Carbon-Sulfur Lyases / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D020869:Gene Expression Profiling / Q000379:methods; D015964:Gene Expression Regulation, Bacterial; D040901:Proteomics / Q000379:methods; D053038:Quorum Sensing / Q000235:genetics / Q000502:physiology*; D018085:Regulon / Q000235:genetics / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D014158:Transcription, Genetic; D041261:Vibrio vulnificus / Q000235:genetics / Q000254:growth & development / Q000378:metabolism / Q000502:physiology*
chemical_list: D001426:Bacterial Proteins; D018797:Cell Cycle Proteins; C118650:SMC protein, Bacteria; D013437:Carbon-Sulfur Lyases; C117753:LuxS protein, Bacteria