1887

Abstract

Previously, the authors have shown that inactivation of , a gene of unknown function upstream of , which encodes a periplasmic disulphide catalyst, results in a global change of gene expression. Among the severely down-regulated genes are , suggesting that the mutant, Sh54, may inefficiently produce the UDP-glucose and UDP-galactose required for LPS synthesis. This paper demonstrates that LPS synthesis in Sh54 is impaired. As a result, Sh54 is unable to polymerize host cell actin, due to aberrant localization of IcsA, or to cause keratoconjunctivitis in guinea pigs. Furthermore, Sh54 is more sensitive to some antimicrobial agents, and exhibits epithelial cytotoxicity characteristic of neither wild-type nor mutants. Supplying restores LPS synthesis and corrects all the defects. Hence, it is clear that the gene is important not only in regulating global gene expression, as shown previously, but also in virulence through LPS synthesis via regulating the expression of the operon.

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2004-04-01
2019-10-15
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References

  1. Belin, P. & Boquet, P. L. ( 1994; ). The Escherichia coli dsbA gene is partly transcribed from the promoter of a weakly expressed upstream gene. Microbiology 140, 3337–3348.[CrossRef]
    [Google Scholar]
  2. Cossart, P. ( 2000; ). Actin-based motility of pathogens: the Arp2/3 complex is a central player. Cell Microbiol 2, 195–205.[CrossRef]
    [Google Scholar]
  3. Genevaux, P., Bauda, P., DuBow, M. S. & Oudega, B. ( 1999; ). Identification of Tn10 insertions in the rfaG, rfaP, and galU genes involved in lipopolysaccharide core biosynthesis that affect Escherichia coli adhesion. Arch Microbiol 172, 1–8.[CrossRef]
    [Google Scholar]
  4. Hitchcock, P. J. & Brown, T. M. ( 1983; ). Morphological heterogeneity among Salmonella lipopolysaccharide chemotypes in silver-stained polyacrylamide gels. J Bacteriol 154, 269–277.
    [Google Scholar]
  5. Hung, D. L., Raivio, T. L., Jones, C. H., Silhavy, T. J. & Hultgren, S. J. ( 2001; ). Cpx signaling pathway monitors biogenesis and affects assembly and expression of P pili. EMBO J 20, 1508–1518.[CrossRef]
    [Google Scholar]
  6. Köhler, H., Rodrigues, S. P. & McCormick, B. A. ( 2002; ). Shigella flexneri interactions with the basolateral membrane domain of polarized model intestinal epithelium: role of lipopolysaccharide in cell invasion and in activation of the mitogen-activated protein kinase ERK. Infect Immun 70, 1150–1158.[CrossRef]
    [Google Scholar]
  7. Li, M. S., Kroll, J. S. & Yu, J. ( 2001; ). Influence of the yihE gene of Shigella flexneri on global gene expression: on analysis using DNA arrays. Biochem Biophys Res Commun 288, 91–100.[CrossRef]
    [Google Scholar]
  8. Nesper, J., Lauriano, C. M., Klose, K. E., Kapfhammer, D., Kraiss, A. & Reidl, J. ( 2001; ). Characterization of Vibrio cholerae O1 El Tor galU and galE mutants: influence on lipopolysaccharide structure, colonization, and biofilm formation. Infect Immun 69, 435–445.[CrossRef]
    [Google Scholar]
  9. Oaks, E. V., Wingfield, M. E. & Formal, S. B. ( 1985; ). Plaque formation by virulent Shigella flexneri. Infect Immun 48, 124–129.
    [Google Scholar]
  10. Okamura, N., Nagai, T., Nakaya, R., Kondo, S., Murakami, M. & Hisatsune, K. ( 1983; ). HeLa cell invasiveness and O antigen of Shigella flexneri as separate and prerequisite attributes of virulence to evoke keratoconjunctivitis in guinea pigs. Infect Immun 39, 505–513.
    [Google Scholar]
  11. Otto, K. & Silhavy, T. J. ( 2002; ). Surface sensing and adhesion of Escherichia coli controlled by the Cpx-signaling pathway. Proc Natl Acad Sci U S A 99, 2287–2292.[CrossRef]
    [Google Scholar]
  12. Raivio, T. L. & Silhavy, T. J. ( 1999; ). The sigmaE and Cpx regulatory pathways: overlapping but distinct envelope stress responses. Curr Opin Microbiol 2, 159–165.[CrossRef]
    [Google Scholar]
  13. Rajakumar, K., Jost, B. H., Sasakawa, C., Okada, N., Yoshikawa, M. & Adler, B. ( 1994; ). Nucleotide sequence of the rhamnose biosynthetic operon of Shigella flexneri 2a and role of lipopolysaccharide in virulence. J Bacteriol 176, 2362–2373.
    [Google Scholar]
  14. Sandlin, R. C., Lampel, K. A., Keasler, S. P., Goldberg, M. B., Stolzer, A. L. & Maurelli, A. T. ( 1995; ). Avirulence of rough mutants of Shigella flexneri: requirement of O antigen for correct unipolar localization of IcsA in the bacterial outer membrane. Infect Immun 63, 229–237.
    [Google Scholar]
  15. Sansonetti, P. J. ( 2001; ). Microbes and microbial toxins: paradigms for microbial-mucosal interactions. III. Shigellosis: from symptoms to molecular pathogenesis. Am J Physiol Gastrointest Liv Physiol 280, G319–G323.
    [Google Scholar]
  16. Sansonetti, P. J., Ryter, A., Clerc, P., Maurelli, A. T. & Mounier, J. ( 1986; ). Multiplication of Shigella flexneri within HeLa cells: lysis of the phagocytic vacuole and plasmid-mediated contact hemolysis. Infect Immun 51, 461–469.
    [Google Scholar]
  17. Schagger, H. & von Jagow, G. ( 1987; ). Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem 166, 368–379.[CrossRef]
    [Google Scholar]
  18. Tsai, C. M. & Frasch, C. E. ( 1982; ). A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal Biochem 119, 115–119.[CrossRef]
    [Google Scholar]
  19. Vaara, M. ( 1992; ). Agents that increase the permeability of outer membrane. Microbiol Rev 56, 395–411.
    [Google Scholar]
  20. Yu, J. ( 1998; ). Inactivation of DsbA, but not DsbC and DsbD, affects the intracellular survival and virulence of Shigella flexneri. Infect Immun 66, 3909–3917.
    [Google Scholar]
  21. Yu, J., Edwards-Jones, B., Neyrolles, O. & Kroll, J. S. ( 2000; ). Key role for DsbA in cell-to-cell spread of Shigella flexneri, permitting secretion of Ipa proteins into interepithelial protrusions. Infect Immun 68, 6449–6456.[CrossRef]
    [Google Scholar]
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