Browse > Article

Identification and Characterization of the Vibrio vulnificus rtxA Essential for Cytotoxicity in vitro and Virulence in Mice  

Lee, Jeong-Hyun (Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University)
Kim, Myung-Won (Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University)
Kim, Byoung-Sik (Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University)
Kim, Seung-Min (Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University)
Lee, Byung-Cheol (School of Life Sciences and Biotechnology, Korea University)
Kim, Tae-Sung (School of Life Sciences and Biotechnology, Korea University)
Choi, Sang-Ho (Department of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University)
Publication Information
Journal of Microbiology / v.45, no.2, 2007 , pp. 146-152 More about this Journal
Abstract
A mutant exhibiting decreased cytotoxic activity toward INT-407 intestinal epithelial cells and carrying a mutation in the rtx gene cluster that consists of rtxCA and rtxBDE operons was screened from a library of V. vulnificus mutants. The functions of the rtxA gene, assessed by constructing an isogenic mutant and evaluating its phenotypic changes, demonstrated that RtxA is essential for the virulence of V. vulnificus in mice as well as in tissue cultures.
Keywords
Vibrio vulnificus; RtxA; cytotoxicity; virulence;
Citations & Related Records
Times Cited By KSCI : 4  (Citation Analysis)
Times Cited By Web Of Science : 54  (Related Records In Web of Science)
Times Cited By SCOPUS : 49
연도 인용수 순위
1 Bradford, M.M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principles of protein-dye binding. Anal. Biochem. 72, 248-254   DOI   ScienceOn
2 Gulig, P.A., K.L. Bourdage, and A.M. Starks. 2005. Molecular pathogenesis of Vibrio vulnificus. J. Microbiol. 43, 118-131   과학기술학회마을
3 Jeong, K.C., H.S. Jeong, J.H. Rhee, S.E. Lee, S.S. Chung, A.M. Starks, G.M. Escudero, P.A. Gulig, and S.H. Choi. 2000. Construction and phenotypic evaluation of Vibrio vulnificus vvpE mutant for elastolytic protease. Infect. Immun. 68, 5096-5106   DOI
4 Kim, H.J., J.H. Lee, J.E. Rhee, H.S. Jeong, H.K. Choi, H.J. Chung, S. Ryu, and S.H. Choi. 2002. Identification and functional analysis of the putAP genes encoding Vibrio vulnificus proline dehydrogenase and proline permease. J. Microbiol. Biotechnol. 12, 318-326
5 Lee, B.C., S.H. Choi, and T.S. Kim. 2003a. Application of sulforhodamine B assay for determining cytotoxicity of Vibrio vulnificus against human intestinal cells. J. Microbiol. Biotechnol. 14, 350-355
6 Lee, J.H., N.Y. Park, S.J. Park, and S.H. Choib. 2003b. Identification and characterization of the Vibrio vulnificus phosphomannomutase gene. J. Microbiol. Biotechnol. 13, 149-154
7 Miller, V.L. and J.J. Mekalanos. 1988. A novel suicide vector and its use in construction of insertion mutations: osmoregulation of outer membrane proteins and virulence determinants in Vibrio cholerae requires toxR. J. Bacteriol. 170, 2575-2583   DOI   PUBMED
8 Park, N.Y., J.H. Lee, B.C. Lee, T.S. Kim, and S.H. Choi. 2006a. Identification and characterization of the wbpO gene essential for lipopolysaccharide synthesis in Vibrio vulnificus. J. Microbiol. Biotechnol. 16, 808-816   과학기술학회마을
9 Park, N.Y., J.H. Lee, M.W. Kim, H.G. Jeong, B.C. Lee, T.S. Kim, and S.H. Choi. 2006b. Identificatioin of Vibrio vulnificus wbpP gene and evaluation of its role in virulence. Infect. Immun. 74, 721-728   DOI   ScienceOn
10 Sheahan, K.L., C.L. Cordero, and K.J. Satchell. 2004. Identification of a domain within the multifunctional Vibrio cholerae RTX toxin that covalently cross-links actin. Proc. Natl. Acad. Sci. USA. 101, 9798-9803
11 Valdivieso-Garcia, A., R.C. Clarke, K. Rahn, A. Durette, D.L. MacLeod, and C.L. Gyles. 1993. Neutral red assay for measurement of quantitative Vero cell cytotoxicity. Appl. Environ. Microbiol. 59, 1981-1983   PUBMED
12 Mekalanos, J.J. 1992. Environmental signals controlling expression of virulence determinants in bacteria. J. Bacteriol. 174, 1-7   DOI   PUBMED
13 Lin, W., K.J. Fullner, R. Clayton, J.A. Sexton, M.B. Rogers, K.E. Calia, S.B. Calderwood, C. Fraser, and J. Mekalanos. 1999. Identification of a Vibrio cholerae RTX toxin gene cluster that is tightly linked to the cholera toxin prophage. Proc. Natl. Acad. Sci. USA. 96, 1071-1076
14 Park, S.G., C.W. Kho, S. Cho, D.H. Lee, S.H. Kim, and B.C. Park. 2002. A functional proteomic analysis of secreted fibrinolytic enzymes from Bacillus subtilis 168 using a combined method of two-dimensional gel electrophoresis and zymography. Proteomics 2, 206-211   DOI   ScienceOn
15 Lee, J.H., S.Y. Jeong, and S.H. Choi. 2006. Regulatory characteristics of the Vibrio vulnificus putAP operon encoding proline dehydrogenase and proline permease. J. Microbiol. Biotechnol. 16, 1285-1291   과학기술학회마을
16 Lee, J.H. and S.H. Choi. 2006. Coactivation of Vibrio vulnificus putAP operon by cAMP receptor protein and PutR through cooperative binding to overlapping sites. Mol. Microbiol. 60, 513-524   DOI   ScienceOn
17 Milton, D.L., R. O'Toole, P. Horstedt, and H. Wolf-Watz. 1996. Flagellin A is essential for the virulence of Vibrio anguillarum. J. Bacteriol. 178, 1310-1319   DOI   PUBMED
18 Boardman, B.K. and K.J. Satchell. 2004. Vibrio cholerae strains with mutations in an atypical type I secretion system accumulate RTX toxin intracellularly. J. Bacteriol. 186, 8137-8143   DOI   ScienceOn
19 Lee, S.H. and A. Camilli. 2000. Novel approaches to monitor bacterial gene expression in infected tissue and host. Curr. Opin. Microbiol. 3, 97-101   DOI   ScienceOn
20 Mahan, M.J., J.M. Slauch, and J.J. Mekalanos. 1993. Selection of bacterial virulence genes that are specifically induced in host tissues. Science 259, 686-688   DOI
21 Rhee, J.E., J.H. Rhee, P.Y. Ryu, and S.H. Choi. 2002. Identification of the cadBA operon from Vibrio vulnificus and its influence on survival to acid stress. FEMS Microbiol. Lett. 208, 245-2512   DOI
22 Lally, E.T., R.B. Hill, I.R. Kieba, and J. Korostoff. 1999. The interaction between RTX toxins and target cells. Trends Microbiol. 7, 356-361   DOI   ScienceOn
23 de Lorenzo, V., M. Herrero, U. Jakubzik, and K.N. Timmis. 1990. Mini-Tn5 transposon derivatives for insertion mutagenesis, promoter probing, and chromosomal insertion of cloned DNA in Gram-negative eubacteria. J. Bacteriol. 172, 6568-6572   DOI   PUBMED
24 Ju, H.M., I.G. Hwang, G.J. Woo, T.S. Kim, and S.H. Choi. 2005. Identification of the Vibrio vulnificus fexA gene and evaluation of its influence on virulence. J. Microbiol. Biotechnol. 15, 1337-1345   과학기술학회마을
25 Laemmli, U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacterophage T4. Nature 227, 680-685   DOI   ScienceOn
26 Strom, M. and R.N. Paranjpye. 2000. Epidemiology and pathogenesis of Vibrio vulnificus. Microbes Infect. 2, 177-188   DOI   ScienceOn
27 Hensel, M., J.E. Shea, C. Gleeson, M.D. Jones, E. Dalton, and D.W. Holden. 1995. Simultaneous identification of bacterial virulence genes by negative selection. Science 269, 400-403   DOI
28 Linkous, D.A. and J.D. Oliver. 1999. Pathogenesis of Vibrio vulnificus. FEMS Microbiol. Lett. 174, 207-214   DOI
29 Oka, A., H. Sugisaki, and M. Takanami. 1981. Nucleotide sequence of the kanamycin resistance transposon Tn903. J. Mol. Biol. 147, 217-226   DOI
30 Jeong, H.S., M.H. Lee, K.H. Lee, S.J. Park, and S.H. Choi. 2003. SmcR and cyclic AMP receptor protein coactivate Vibrio vulnificus vvpE encoding elastase through the RpoS-dependent promoter in a synergistic manner. J. Biol. Chem. 278, 45072-45081   DOI   ScienceOn
31 Kim, Y.R., S.E. Lee, C.M. Kim, S.Y. Kim, E.K. Shin, D.H. Shin, S.S. Chung, H.E. Choy, A. Progulske-Fox, J.D. Hillman, M. Handfield, and J.H. Rhee. 2003. Characterization and pathogenic significance of Vibrio vulnificus antigens preferentially expressed in septicemic patients. Infect. Immun. 71, 5461-5471   DOI
32 Rhee, J.E., H.M. Ju, U. Park, BC. Park, and S.H. Choi. 2004. Identification of the Vibrio vulnificus cadC and evaluation of its role in acid tolerance. J. Microbiol. Biotechnol. 14, 1093-1098
33 Murphy, K.C., K.G. Campellone, and A.R. Poteete. 2000. PCRmediated gene replacement in Escherichia coli. Gene 246, 321-330   DOI   ScienceOn
34 Reed, L.J. and H. Muench. 1938. A simple method of estimating fifty percent endpoints. Am. J. Hyg. 27, 439-497
35 Fullner, K.J. and J.J. Mekalanos. 2000. In vivo covalent cross-linking of cellular actin by the Vibrio cholerae RTX toxin. EMBO J. 19, 5315-5323   DOI   ScienceOn
36 Kachlany, S.C., D.H. Fine, and D.H. Figurski. 2002. Purification of secreted leukotoxin (LtxA) from Actinobacillus actinomycetemcomitans. Protein Expr. Purif. 25, 465-471   DOI   ScienceOn