Proteomic Identification and Characterization of Vibrio vulnificus Proteins Induced upon Exposure to INT-407 Intestinal Epithelial Cells

  • Oh, Man-Hwan (National Research Laboratory of Molecular Microbiology and Toxicology, Department of Agricultural Biotechnology, and Center for Agricultural Biomaterials, Seoul National University) ;
  • Jeong, Hee-Gon (National Research Laboratory of Molecular Microbiology and Toxicology, Department of Agricultural Biotechnology, and Center for Agricultural Biomaterials, Seoul National University) ;
  • Choi, Sang-Ho (National Research Laboratory of Molecular Microbiology and Toxicology, Department of Agricultural Biotechnology, and Center for Agricultural Biomaterials, Seoul National University)
  • Published : 2008.05.31

Abstract

Proteomic analysis led to identification of the proteins of Vibrio vulnificus that were induced upon exposure to INT-407 cells, and 7 of which belong to the functional categories such as amino acid transport/metabolism, nucleotide transport/metabolism, posttranslational modification/protein turnover/chaperones, and translation. Among the genes encoding the host-induced proteins, disruption of purH, trpD, tsaA, and groEL2 resulted in reduced cytotoxicity. The purH, trpD, and tsuA mutants showed impaired growth in the INT-407 lysate; however, the growth rate of the groEL2 mutant was not significantly changed, indicating that the possible roles of the host-induced proteins in the virulence of V. vulnificus are rather versatile.

Keywords

References

  1. Abshire, K. Z. and F. C. Neidhardt. 1993. Analysis of proteins synthesized by Salmonella typhimurium during growth within a host macrophage. J. Bacteriol. 175: 3734-3743 https://doi.org/10.1128/jb.175.12.3734-3743.1993
  2. Bergman, N. H., E. C. Anderson, E. E. Swenson, B. K. Janes, N. Fisher, M. M. Niemeyer, A. D. Miyoshi, and P. C. Hanna. 2007. Transcriptional profiling of Bacillus anthracis during infection of host macrophages. Infect. Immun. 75: 3434-3444 https://doi.org/10.1128/IAI.01345-06
  3. Chiang, S. L., J. J. Mekalanos, and D. W. Holden. 1999. In vivo genetic analysis of bacterial virulence. Annu. Rev. Microbiol. 53: 129-154 https://doi.org/10.1146/annurev.micro.53.1.129
  4. De Finlay, B. B., F. Heffron, and S. Falkow. 1989. Epithelial cell surfaces induce Salmonella proteins required for bacterial adherence and invasion. Science 243: 940-943 https://doi.org/10.1126/science.2919285
  5. Faucher, S. P., S. Porwollik, C. M. Dozois, M. McClelland, and F. Daigle. 2006. Transcriptome of Salmonella enterica serovar Typhi within macrophages revealed through the selective capture of transcribed sequences. Proc. Natl. Acad. Sci. USA 103: 1906-1911
  6. Goo, S. Y., H. J. Lee, W. H. Kim, K. L. Han, D. K. Park, H. J. Lee, et al. 2006. Identification of OmpU of Vibrio vulnificus as a fibronectin-binding protein and its role in bacterial pathogenesis. Infect. Immun. 74: 5586-5594 https://doi.org/10.1128/IAI.00171-06
  7. Gulig, P. A., K. L. Bourdage, and A. M. Starks. 2005. Molecular pathogenesis of Vibrio vulnificus. J. Microbiol. 43: 118-131
  8. 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 https://doi.org/10.1126/science.7618105
  9. 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 https://doi.org/10.1074/jbc.M308184200
  10. Jeong, K. C., H. S. Jeong, S. E. Lee, S. S. Chung, J. H. Rhee, A. M. Starks, G. M. Escudero, P. A. Gulig, and S. H. Choi. 2000. Construction and phenotypic evaluation of a Vibrio vulnificus vvpE mutant for elastolytic protease. Infect. Immun. 68: 5096-5106 https://doi.org/10.1128/IAI.68.9.5096-5106.2000
  11. Jeong, W., M. K. Cha, and I. H. Kim. 2000. Thioredoxindependent hydroperoxide peroxidase activity of bacterioferritin comigratory protein (BCP) as a new member of the thiolspecific antioxidant protein (TSA)/alkyl hydroperoxide peroxidase C (AhpC) family. J. Biol. Chem. 275: 2924-2930 https://doi.org/10.1074/jbc.275.4.2924
  12. Jones, M. K., E. Warner, and J. D. Oliver. 2008. Survival of and in situ gene expression by Vibrio vulnificus at varying salinities in estuarine environments. Appl. Environ. Microbiol. 74: 182-187 https://doi.org/10.1128/AEM.02436-07
  13. Kim, Y. R., S. E. Lee, C. M. Kim, S. Y. Kim, E. K. Shin, D. H. Shin, et al. 2003. Characterization and pathogenic significance of Vibrio vulnificus antigens preferentially expressed in septicemic patients. Infect. Immun. 71: 5461-5471 https://doi.org/10.1128/IAI.71.10.5461-5471.2003
  14. Konkel, M. E. and W. Jr. Cieplak. 1992. Altered synthetic response of Campylobacter jejuni to cocultivation with human epithelial cells is associated with enhanced internalization. Infect. Immun. 60: 4945-4949
  15. Lee, A. Y., S. G. Park, M. Jang, S. Cho, P. K. Myung, Y. R. Kim, J. H. Rhee, D. H. Lee, and B. C. Park. 2006. Proteomic analysis of pathogenic bacterium Vibrio vulnificus. Proteomics 6: 1284-1289
  16. Lee, J. H., J. E. Rhee, U. R. Park, H. M. Ju, B. C. Lee, T. S. Kim, H. S. Jeong, and S. H. Choi. 2007. Identification and functional analysis of Vibrio vulnificus smcR, a novel global regulator. J. Microbiol. Biotechnol. 17: 325-334
  17. Lee, J. H., M. W. Kim, B. S. Kim, S. M. Kim, B. C. Lee, T. S. Kim, and S. H. Choi. 2007. Identification and characterization of the Vibrio vulnificus rtxA essential for cytotoxicity in vitro and virulence in mice. J. Microbiol. 45: 146-152
  18. Lee, J. Y., J. B. Eun, and S. H. Choi. 1997. Improving detection of Vibrio vulnificus in Octopus variabilis by PCR. J. Food Sci. 62: 179-182 https://doi.org/10.1111/j.1365-2621.1997.tb04395.x
  19. 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
  20. 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 https://doi.org/10.1016/S1369-5274(99)00058-2
  21. Leung, K. and B. Finlay. 1991. Intracellular replication is essential for the virulence of Salmonella typhimurium. Proc. Natl. Acad. Sci. USA 88: 11470-11474
  22. Linkous, D. A. and J. D. Oliver. 1999. Pathogenesis of Vibrio vulnificus. FEMS Microbiol. Lett. 174: 207-214 https://doi.org/10.1111/j.1574-6968.1999.tb13570.x
  23. 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 https://doi.org/10.1126/science.8430319
  24. Mekalanos, J. J. 1992. Environmental signals controlling expression of virulence determinants in bacteria. J. Bacteriol. 174: 1-7 https://doi.org/10.1128/jb.174.1.1-7.1992
  25. 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 https://doi.org/10.1128/jb.170.6.2575-2583.1988
  26. 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 https://doi.org/10.1128/jb.178.5.1310-1319.1996
  27. Oka, A., H. Sugisaki, and M. Takanami. 1981. Nucleotide sequence of the kanamycin resistance transposon Tn903. J. Mol. Biol. 147: 217-226 https://doi.org/10.1016/0022-2836(81)90438-1
  28. Park, N. Y., J. H. Lee, B. C. Lee, T. S. Kim, and S. H. Choi. 2006. Identification and characterization of the wbpO gene essential for lipopolysaccharide synthesis in Vibrio vulnificus. J. Microbiol. Biotechnol. 16: 808-816
  29. Park, N. Y., J. H. Lee, M. W. Kim, H. G. Jeong, B. C. Lee, T. S. Kim, and S. H. Choi. 2006. Identificatioin of Vibrio vulnificus wbpP gene and evaluation of its role in virulence. Infect. Immun. 74: 721-728 https://doi.org/10.1128/IAI.74.1.721-728.2006
  30. 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 https://doi.org/10.1002/1615-9861(200202)2:2<206::AID-PROT206>3.0.CO;2-5
  31. Seaver, L. C. and J. A. Imlay. 2001. Alkyl hydroperoxide reductase is the primary scavenger of endogenous hydrogen peroxide in Escherichia coli. J. Bacteriol. 183: 7173-7181 https://doi.org/10.1128/JB.183.24.7173-7181.2001
  32. Smith, D. A., T. Parish, N. G. Stoker, and G. J. Bancroft. 2001. Characterization of auxotrophic mutants of Mycobacterium tuberculosis and their potential as vaccine candidates. Infect. Immun. 69: 1142-1150 https://doi.org/10.1128/IAI.69.2.1442-1150.2001
  33. Strom, M. and R. N. Paranjpye. 2000. Epidemiology and pathogenesis of Vibrio vulnificus. Microbes Infect. 2: 177-188 https://doi.org/10.1016/S1286-4579(00)00270-7
  34. Torres, A. G. and J. B. Kaper. 2003. Multiple elements controlling adherence of enterohemorrhagic Escherichia coli O157:H7 to HeLa cells. Infect. Immun. 71: 4985-4995 https://doi.org/10.1128/IAI.71.9.4985-4995.2003
  35. Yoon, H. J., S. Y. Lim, S. G. Heu, S. H. Choi, and S. R. Ryu. 2003. Proteome analysis of Salmonella enterica serovar Typhimurium fis mutant. FEMS Microbiol. Lett. 226: 391-396 https://doi.org/10.1016/S0378-1097(03)00641-4
  36. Zugel, U. and S. H. Kaufmann. 1999. Role of heat shock proteins in protection from and pathogenesis of infectious diseases. Clin. Microbiol. Rev. 12: 19-39