Skip to main content
Infection and Immunity logoLink to Infection and Immunity
. 1996 Nov;64(11):4739–4743. doi: 10.1128/iai.64.11.4739-4743.1996

Expression of Salmonella typhimurium rpoS and rpoS-dependent genes in the intracellular environment of eukaryotic cells.

C Y Chen 1, L Eckmann 1, S J Libby 1, F C Fang 1, S Okamoto 1, M F Kagnoff 1, J Fierer 1, D G Guiney 1
PMCID: PMC174440  PMID: 8890234

Abstract

Adaptation to the intracellular environment of host cells is crucial for the pathogenesis of Salmonella infections. The alternative sigma factor RpoS is a global regulator of gene expression during starvation and stress conditions and is required for virulence in Salmonella spp. We have used lacZ reporter fusions to rpoS and rpoS-dependent genes to study rpoS regulation after entry of Salmonella typhimurium into macrophages and epithelial cells. The results demonstrate that expression of an rpoS::lacZ translational fusion increases rapidly in S. typhimurium after phagocytosis. Activity of RpoS also increases after bacterial entry into both macrophages and epithelial cells, as demonstrated by the induction of the rpoS-regulated genes katE and spvB. A control rpoS-independent promoter for neomycin resistance does not show significant induction after cell entry. These results demonstrate that the regulatory system mediated by RpoS in S. typhimurium is activated by the intracellular environment of eukaryotic cells.

Full Text

The Full Text of this article is available as a PDF (232.5 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alpuche Aranda C. M., Swanson J. A., Loomis W. P., Miller S. I. Salmonella typhimurium activates virulence gene transcription within acidified macrophage phagosomes. Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10079–10083. doi: 10.1073/pnas.89.21.10079. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barrow P. A., Simpson J. M., Lovell M. A., Binns M. M. Contribution of Salmonella gallinarum large plasmid toward virulence in fowl typhoid. Infect Immun. 1987 Feb;55(2):388–392. doi: 10.1128/iai.55.2.388-392.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Buchmeier N. A., Heffron F. Induction of Salmonella stress proteins upon infection of macrophages. Science. 1990 May 11;248(4956):730–732. doi: 10.1126/science.1970672. [DOI] [PubMed] [Google Scholar]
  4. Buchmeier N. A., Heffron F. Inhibition of macrophage phagosome-lysosome fusion by Salmonella typhimurium. Infect Immun. 1991 Jul;59(7):2232–2238. doi: 10.1128/iai.59.7.2232-2238.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Buchmeier N. A., Heffron F. Intracellular survival of wild-type Salmonella typhimurium and macrophage-sensitive mutants in diverse populations of macrophages. Infect Immun. 1989 Jan;57(1):1–7. doi: 10.1128/iai.57.1.1-7.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chen C. Y., Buchmeier N. A., Libby S., Fang F. C., Krause M., Guiney D. G. Central regulatory role for the RpoS sigma factor in expression of Salmonella dublin plasmid virulence genes. J Bacteriol. 1995 Sep;177(18):5303–5309. doi: 10.1128/jb.177.18.5303-5309.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chen C. Y., Winans S. C. Controlled expression of the transcriptional activator gene virG in Agrobacterium tumefaciens by using the Escherichia coli lac promoter. J Bacteriol. 1991 Feb;173(3):1139–1144. doi: 10.1128/jb.173.3.1139-1144.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Danbara H., Moriguchi R., Suzuki S., Tamura Y., Kijima M., Oishi K., Matsui H., Abe A., Nakamura M. Effect of 50 kilobase-plasmid, pKDSC50, of Salmonella choleraesuis RF-1 strain on pig septicemia. J Vet Med Sci. 1992 Dec;54(6):1175–1178. doi: 10.1292/jvms.54.1175. [DOI] [PubMed] [Google Scholar]
  9. De Groote M. A., Testerman T., Xu Y., Stauffer G., Fang F. C. Homocysteine antagonism of nitric oxide-related cytostasis in Salmonella typhimurium. Science. 1996 Apr 19;272(5260):414–417. doi: 10.1126/science.272.5260.414. [DOI] [PubMed] [Google Scholar]
  10. Fang F. C., Krause M., Roudier C., Fierer J., Guiney D. G. Growth regulation of a Salmonella plasmid gene essential for virulence. J Bacteriol. 1991 Nov;173(21):6783–6789. doi: 10.1128/jb.173.21.6783-6789.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fang F. C., Libby S. J., Buchmeier N. A., Loewen P. C., Switala J., Harwood J., Guiney D. G. The alternative sigma factor katF (rpoS) regulates Salmonella virulence. Proc Natl Acad Sci U S A. 1992 Dec 15;89(24):11978–11982. doi: 10.1073/pnas.89.24.11978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fields P. I., Swanson R. V., Haidaris C. G., Heffron F. Mutants of Salmonella typhimurium that cannot survive within the macrophage are avirulent. Proc Natl Acad Sci U S A. 1986 Jul;83(14):5189–5193. doi: 10.1073/pnas.83.14.5189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fierer J., Eckmann L., Fang F., Pfeifer C., Finlay B. B., Guiney D. Expression of the Salmonella virulence plasmid gene spvB in cultured macrophages and nonphagocytic cells. Infect Immun. 1993 Dec;61(12):5231–5236. doi: 10.1128/iai.61.12.5231-5236.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fierer J., Hatlen L., Lin J. P., Estrella D., Mihalko P., Yau-Young A. Successful treatment using gentamicin liposomes of Salmonella dublin infections in mice. Antimicrob Agents Chemother. 1990 Feb;34(2):343–348. doi: 10.1128/aac.34.2.343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Garcia-del Portillo F., Zwick M. B., Leung K. Y., Finlay B. B. Salmonella induces the formation of filamentous structures containing lysosomal membrane glycoproteins in epithelial cells. Proc Natl Acad Sci U S A. 1993 Nov 15;90(22):10544–10548. doi: 10.1073/pnas.90.22.10544. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Guiney D. G., Fang F. C., Krause M., Libby S., Buchmeier N. A., Fierer J. Biology and clinical significance of virulence plasmids in Salmonella serovars. Clin Infect Dis. 1995 Oct;21 (Suppl 2):S146–S151. doi: 10.1093/clinids/21.supplement_2.s146. [DOI] [PubMed] [Google Scholar]
  17. Guiney D. G., Libby S., Fang F. C., Krause M., Fierer J. Growth-phase regulation of plasmid virulence genes in Salmonella. Trends Microbiol. 1995 Jul;3(7):275–279. doi: 10.1016/s0966-842x(00)88944-1. [DOI] [PubMed] [Google Scholar]
  18. Gulig P. A., Doyle T. J. The Salmonella typhimurium virulence plasmid increases the growth rate of salmonellae in mice. Infect Immun. 1993 Feb;61(2):504–511. doi: 10.1128/iai.61.2.504-511.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Heffernan E. J., Fierer J., Chikami G., Guiney D. Natural history of oral Salmonella dublin infection in BALB/c mice: effect of an 80-kilobase-pair plasmid on virulence. J Infect Dis. 1987 Jun;155(6):1254–1259. doi: 10.1093/infdis/155.6.1254. [DOI] [PubMed] [Google Scholar]
  20. Jain V. K., Magrath I. T. A chemiluminescent assay for quantitation of beta-galactosidase in the femtogram range: application to quantitation of beta-galactosidase in lacZ-transfected cells. Anal Biochem. 1991 Nov 15;199(1):119–124. doi: 10.1016/0003-2697(91)90278-2. [DOI] [PubMed] [Google Scholar]
  21. Jones B. D., Ghori N., Falkow S. Salmonella typhimurium initiates murine infection by penetrating and destroying the specialized epithelial M cells of the Peyer's patches. J Exp Med. 1994 Jul 1;180(1):15–23. doi: 10.1084/jem.180.1.15. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lange R., Hengge-Aronis R. The cellular concentration of the sigma S subunit of RNA polymerase in Escherichia coli is controlled at the levels of transcription, translation, and protein stability. Genes Dev. 1994 Jul 1;8(13):1600–1612. doi: 10.1101/gad.8.13.1600. [DOI] [PubMed] [Google Scholar]
  23. Loewen P. C., Hengge-Aronis R. The role of the sigma factor sigma S (KatF) in bacterial global regulation. Annu Rev Microbiol. 1994;48:53–80. doi: 10.1146/annurev.mi.48.100194.000413. [DOI] [PubMed] [Google Scholar]
  24. Loewen P. C., Triggs B. L. Genetic mapping of katF, a locus that with katE affects the synthesis of a second catalase species in Escherichia coli. J Bacteriol. 1984 Nov;160(2):668–675. doi: 10.1128/jb.160.2.668-675.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Loewen P. C., von Ossowski I., Switala J., Mulvey M. R. KatF (sigma S) synthesis in Escherichia coli is subject to posttranscriptional regulation. J Bacteriol. 1993 Apr;175(7):2150–2153. doi: 10.1128/jb.175.7.2150-2153.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Maw J., Meynell G. G. The true division and death rates of Salmonella typhimurium in the mouse spleen determined with superinfecting phage P22. Br J Exp Pathol. 1968 Dec;49(6):597–613. [PMC free article] [PubMed] [Google Scholar]
  27. McCann M. P., Fraley C. D., Matin A. The putative sigma factor KatF is regulated posttranscriptionally during carbon starvation. J Bacteriol. 1993 Apr;175(7):2143–2149. doi: 10.1128/jb.175.7.2143-2149.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Miller V. L., Mekalanos J. J. 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. 1988 Jun;170(6):2575–2583. doi: 10.1128/jb.170.6.2575-2583.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Muffler A., Traulsen D. D., Lange R., Hengge-Aronis R. Posttranscriptional osmotic regulation of the sigma(s) subunit of RNA polymerase in Escherichia coli. J Bacteriol. 1996 Mar;178(6):1607–1613. doi: 10.1128/jb.178.6.1607-1613.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Mulvey M. R., Switala J., Borys A., Loewen P. C. Regulation of transcription of katE and katF in Escherichia coli. J Bacteriol. 1990 Dec;172(12):6713–6720. doi: 10.1128/jb.172.12.6713-6720.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Notley L., Ferenci T. Induction of RpoS-dependent functions in glucose-limited continuous culture: what level of nutrient limitation induces the stationary phase of Escherichia coli? J Bacteriol. 1996 Mar;178(5):1465–1468. doi: 10.1128/jb.178.5.1465-1468.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Rhen M., Riikonen P., Taira S. Transcriptional regulation of Salmonella enterica virulence plasmid genes in cultured macrophages. Mol Microbiol. 1993 Oct;10(1):45–56. doi: 10.1111/j.1365-2958.1993.tb00902.x. [DOI] [PubMed] [Google Scholar]
  33. Schweder T., Lee K. H., Lomovskaya O., Matin A. Regulation of Escherichia coli starvation sigma factor (sigma s) by ClpXP protease. J Bacteriol. 1996 Jan;178(2):470–476. doi: 10.1128/jb.178.2.470-476.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Takayanagi Y., Tanaka K., Takahashi H. Structure of the 5' upstream region and the regulation of the rpoS gene of Escherichia coli. Mol Gen Genet. 1994 Jun 3;243(5):525–531. doi: 10.1007/BF00284200. [DOI] [PubMed] [Google Scholar]
  35. Wallis T. S., Paulin S. M., Plested J. S., Watson P. R., Jones P. W. The Salmonella dublin virulence plasmid mediates systemic but not enteric phases of salmonellosis in cattle. Infect Immun. 1995 Jul;63(7):2755–2761. doi: 10.1128/iai.63.7.2755-2761.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES