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

Authors

  • Na-Ri Shin,

    1. Department of Infectious Diseases, BK21 for Veterinary Science and KRF Zoonotic Priority Research Institute, College of Veterinary Medicine, Seoul National University, Korea
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  • Deog Yong Lee,

    1. Department of Infectious Diseases, BK21 for Veterinary Science and KRF Zoonotic Priority Research Institute, College of Veterinary Medicine, Seoul National University, Korea
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  • Han Sang Yoo

    1. Department of Infectious Diseases, BK21 for Veterinary Science and KRF Zoonotic Priority Research Institute, College of Veterinary Medicine, Seoul National University, Korea
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  • Present address: Na-Ri Shin, Center for Infectious Diseases, Division of Biodefense Research, Korea National Institute for Health, Seoul, 122-701 Korea.

  • Editor: Kai Man Kam

Correspondence: Han Sang Yoo, Department of Infectious Diseases, College of Veterinary Medicine, Seoul National University, Seoul 151-742, Korea. Tel.: +82 2 880 1263; fax: +82 2 874 2738; e-mail: yoohs@snu.ac.kr

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, VvSRΔZ, and a luxS–smcR double mutant, VvSRΔZSR, 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.

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