1887

Abstract

A two-component regulatory system, , plays an important role in the pH-dependent regulation of , a global activator for virulence determinants including invasion genes, in . The authors examined whether the homologues have some function in the expression of serovar Typhimurium invasion genes via the regulation of , an activator for these genes. In a mutant, the expression level was reduced to less than 10 % of that in the parent strain at pH 6·0. This mutant strain also showed undetectable synthesis of an invasion gene product, SipC, at pH 6·0 and reduced cell invasion capacity – as low as 20 % of that of the parent. In this mutant, the reduction in expression was much less marked at pH 8·0 than at pH 6·0 – no less than 50 % of that in the parent, and no significant reduction was observed in either SipC synthesis or cell invasion rate, compared to the parent. Unexpectedly, a mutant strain and the parent showed no apparent difference in all three characteristics described above at either pH. These results indicate that in , the sensor kinase CpxA activates , and consequently, invasion genes and cell invasion capacity at pH 6·0. At pH 8·0, however, CpxA does not seem to have a large role in activation of these factors. Further, the results show that this CpxA-mediated activation does not require its putative cognate response regulator, CpxR. This suggests that CpxA may interact with regulator(s) other than CpxR to achieve activation at low pH.

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2003-10-01
2020-05-26
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References

  1. Altier C., Suyemoto M., Ruiz A. I., Burnham K. D., Maurer R.. 2000a; Characterization of two novel regulatory genes affecting Salmonella invasion gene expression. Mol Microbiol35:635–646
    [Google Scholar]
  2. Altier C., Suyemoto M., Lawton S. D.. 2000b; Regulation of Salmonella enterica Serovar Typhimurium invasion genes by csrA . Infect Immun68:6790–6797
    [Google Scholar]
  3. Bajaj V., Hwang C., Lee C. A.. 1995; hilA is a novel ompR/toxR family member that activates the expression of Salmonella typhimurium invasion genes. Mol Microbiol18:715–727
    [Google Scholar]
  4. Bajaj V., Lucas R. L., Hwang C., Lee C. A.. 1996; Co-ordinate regulation of Salmonella typhimurium invasion genes by environmental and regulatory factors is mediated by control of hilA expression. Mol Microbiol22:703–714
    [Google Scholar]
  5. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W.. 1977; Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene2:95–113
    [Google Scholar]
  6. Danese P. N., Silhavy T. J.. 1998; CpxP, a stress-combative member of the Cpx regulon. J Bacteriol180:831–839
    [Google Scholar]
  7. Danese P. N., Snyder W. B., Cosma C. L., Davis L. J. B., Silhavy T. J.. 1995; The Cpx two-component signal transduction pathway of Escherichia coli regulates transcription of the gene specifying the stress-inducible periplasmic protease, DegP. Genes Dev9:387–398
    [Google Scholar]
  8. Darwin K. H., Miller V. L.. 1999; Molecular basis of the interaction of Salmonella with the intestinal mucosa. Clin Microbiol Rev12:405–428
    [Google Scholar]
  9. Dong J., Iuchi S., Kwan H.-S., Lu Z., Lin E. C. C.. 1993; The deduced amino-acid sequence of the cloned cpxR gene suggests the protein is the cognate regulator for the membrane sensor, CpxA, in a two-component signal transduction system of Escherichia coli . Gene136:227–230
    [Google Scholar]
  10. Dorman C. J., Porter M. E.. 1998; The Shigella virulence gene regulatory cascade: a paradigm of bacterial gene control mechanisms. Mol Microbiol29:677–684
    [Google Scholar]
  11. Eichelberg K., Galan J. E.. 1999; Differential regulation of Salmonella typhimurium Type III secreted proteins by Pathogenicity Island 1 (SPI-1)-encoded transcriptional activators InvF and HilA. Infect Immun67:4099–4105
    [Google Scholar]
  12. Fahlen T. F., Mathur N., Jones B. D.. 2000; Identification and characterization of mutants with increased expression of hilA , the invasion gene transcriptional activator of Salmonella typhimurium . FEMS Immunol Med Microbiol28:25–35
    [Google Scholar]
  13. Galan J. E.. 1996; Molecular genetic bases of Salmonella entry into host cells. Mol Microbiol20:263–271
    [Google Scholar]
  14. Huang X.-Z., Tall B., Schwan W. R., Kopecko D. J.. 1998; Physical limitations on Salmonella typhi entry into cultured human intestinal epithelial cells. Infect Immun66:2928–2937
    [Google Scholar]
  15. Iyoda S., Kamidoi T., Hirose K., Kutsukake K., Watanabe H.. 2001; A flagellar gene fliZ regulates the expression of invasion genes and virulence phenotype in Salmonella enterica Serovar Typhimurium. Microb Pathog30:81–90
    [Google Scholar]
  16. Johnson K., Charles I., Dougan G., Pickard D., O'Gaora P., Costa G., Ali T., Miller I., Hormaeche C.. 1991; The role of a stress-response protein in Salmonella typhimurium virulence. Mol Microbiol5:401–407
    [Google Scholar]
  17. Johnston C., Pegues D. A., Hueck C. J., Lee C. A., Miller S. I.. 1996; Transcriptional activation of Salmonella typhimurium invasion genes by a member of the phosphorylated response-regulator superfamily. Mol Microbiol22:715–727
    [Google Scholar]
  18. Kutsukake K., Iyoda S., Ohnishi K., Iino T.. 1994; Genetic and molecular analyses of the interaction between the flagellum-specific sigma and anti-sigma factors in Salmonella typhimurium . EMBO J13:4568–4576
    [Google Scholar]
  19. Leclerc G. J., Tartera C., Metcalf E. S.. 1998; Environmental regulation of Salmonella typhi invasion-defective mutants. Infect Immun66:682–691
    [Google Scholar]
  20. Lucas R. L., Lee C. A.. 2000; Unravelling the mysteries of virulence gene regulation in Salmonella typhimurium . Mol Microbiol36:1024–1033
    [Google Scholar]
  21. Lucas R. L., Lee C. A.. 2001; Roles of hilC and hilD in regulation of hilA expression in Salmonella enterica Serovar Typhimurium. J Bacteriol183:2733–2745
    [Google Scholar]
  22. Lucas R. L., Lostroh C. P., DiRusso C. C., Spector M. P., Wanner B. L., Lee C. A.. 2000; Multiple factors independently regulate hilA and invasion gene expression in Salmonella enterica Serovar Typhimurium. J Bacteriol182:1872–1882
    [Google Scholar]
  23. Maniatis T., Fritsch E. F., Sambrook J.. 1982; Molecular Cloning: a Laboratory Manual Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  24. McClelland M., Sanderson K. E., Spieth J.. 23 other authors 2001; Complete genome sequence of Salmonella enterica serovar Typhimurium LT2. Nature413:852–856
    [Google Scholar]
  25. Miller J. H.. 1972; Experiments in Molecular Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  26. Nakayama S., Watanabe H.. 1995; Involvement of cpxA , a sensor of a two-component regulatory sysytem, in the pH-dependent regulation of expression of Shigella sonnei virF gene. J Bacteriol177:5062–5069
    [Google Scholar]
  27. Nakayama S., Watanabe H.. 1998; Identification of cpxR as a positive regulator essential for expression of the Shigella sonnei virF gene. J Bacteriol180:3522–3528
    [Google Scholar]
  28. Ohmori H., Saitoh M., Yasuda T., Nagata T., Fujii T., Wachi M., Nagai K.. 1995; The pcsA gene is identical to dinD in Escherichia coli . J Bacteriol177:156–165
    [Google Scholar]
  29. Oka A., Sugisaki H., Takanami M.. 1981; Nucleotide sequence of the kanamycin resistance transposon Tn 903 . J Mol Biol147:217–226
    [Google Scholar]
  30. Pegues D. A., Hantman M. J., Behlau I., Miller S. I.. 1995; PhoP/PhoQ transcriptional repression of Salmonella typhimurium invasion genes: evidence for a role in protein secretion. Mol Microbiol17:169–181
    [Google Scholar]
  31. Schechter L. M., Damrauer S. M., Lee C. A.. 1999; Two AraC/XylS family members can independently counteract the effect of repressing sequences upstream of the hilA promoter. Mol Microbiol32:629–642
    [Google Scholar]
  32. Schmieger H.. 1972; Phage P22 -mutants with increased or decreased transduction abilities. Mol Gen Genet119:75–88
    [Google Scholar]
  33. Shapira S. K., Chou J., Richaud F. V., Casadaban M. J.. 1983; New versatile plasmid vectors for expression of hybrid proteins coded by a cloned gene fused to lacZ gene sequences encoding an enzymatically active carboxy-terminal portion of β -galactosidase. Gene25:71–82
    [Google Scholar]
  34. Takeshita S., Sato M., Toba M., Masahashi W., Hashimoto-Gotoh T.. 1987; High-copy-number and low-copy-number plasmid vectors for lacZ α -complementation and chloramphenicol- or kanamycin-resistance selection. Gene61:63–74
    [Google Scholar]
  35. Weber R. F., Silverman P. M.. 1988; The Cpx proteins of Escherichia coli K12: structure of the CpxA polypeptide as an inner membrane component. J Mol Biol203:467–478
    [Google Scholar]
  36. Wilson R. L., Libby S. J., Freet A. M., Boddicker J. D., Fahlen T. F., Jones B. D.. 2001; Fis, a DNA nucleoid-associated protein, is involved in Salmonella typhimurium SPI-1 invasion gene expression. Mol Microbiol39:79–88
    [Google Scholar]
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