Identification of quorum sensing-related regulons in Vibrio vulnificus by two-dimensional gel electrophoresis and differentially displayed reverse transcriptase PCR.
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.
Shin
Na-Ri
NR
Department of Infectious Diseases, BK21 for Veterinary Science and KRF Zoonotic Priority Research Institute, College of Veterinary Medicine, Seoul National University, Korea.
Lee
Deog Yong
DY
Yoo
Han Sang
HS
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*
D001426:Bacterial Proteins; D018797:Cell Cycle Proteins; C118650:SMC protein, Bacteria; D013437:Carbon-Sulfur Lyases; C117753:LuxS protein, Bacteria