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. 1997 May;65(5):1814–1823. doi: 10.1128/iai.65.5.1814-1823.1997

Role of sigma factor RpoS in initial stages of Salmonella typhimurium infection.

C A Nickerson 1, R Curtiss 3rd 1
PMCID: PMC175223  PMID: 9125566

Abstract

The sigma factor RpoS mediates the stationary-phase expression of a large group of genes, including those involved in resistance to a variety of environmental stresses, such as starvation, oxidation, and low pH. In addition, RpoS has been shown to regulate Salmonella virulence. In Salmonella typhimurium, RpoS controls the expression of the Salmonella plasmid virulence (spv) genes, which are required for systemic infection. However, the mechanism by which RpoS affects the pathogenicity of Salmonella remains incompletely defined. In this study, we focused on the ability of rpoS to affect the early stages of the infection process of S. typhimurium. An rpoS mutant of S. typhimurium exhibited wild-type abilities to attach to and invade Int-407 cells and J774 macrophage-like cells. In addition, rpoS did not affect the intracellular survival of S. typhimurium in either J774 macrophage-like cells or rat bone marrow-derived macrophages. However, the rpoS mutant demonstrated a decreased ability to colonize murine Peyer's patches after oral inoculation than its wild-type virulent parent strain showed. In addition, virulence plasmid-cured derivatives of the rpoS mutant were recovered in lower numbers from murine Peyer's patches than were plasmid-cured derivatives of the isogenic wild-type S. typhimurium. This indicates that RpoS regulation of chromosomally encoded genes is important for colonization of the gut-associated lymphoid tissue (GALT) by S. typhimurium. Microscopic analysis of histological sections taken from Peyer's patches after peroral infection of mice showed that, unlike its wild-type virulent parent strain, the isogenic rpoS mutant did not destroy the follicle-associated epithelium of the GALT. Furthermore, the rpoS mutant demonstrated a decreased ability to adhere to histological sections of murine Peyer's patches than its wild-type parent showed. Our data provide evidence for a role of RpoS in the interaction of Salmonella with cells of the GALT, specifically the Peyer's patches. This implicates the involvement of rpoS in the initial stages of systemic infection by Salmonella as opposed to infection leading to gastroenteritis.

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Selected References

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  1. Baird G. D., Manning E. J., Jones P. W. Evidence for related virulence sequences in plasmids of Salmonella dublin and Salmonella typhimurium. J Gen Microbiol. 1985 Jul;131(7):1815–1823. doi: 10.1099/00221287-131-7-1815. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Bäumler A. J., Tsolis R. M., Heffron F. The lpf fimbrial operon mediates adhesion of Salmonella typhimurium to murine Peyer's patches. Proc Natl Acad Sci U S A. 1996 Jan 9;93(1):279–283. doi: 10.1073/pnas.93.1.279. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Caldwell A. L., Gulig P. A. The Salmonella typhimurium virulence plasmid encodes a positive regulator of a plasmid-encoded virulence gene. J Bacteriol. 1991 Nov;173(22):7176–7185. doi: 10.1128/jb.173.22.7176-7185.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Carter P. B., Collins F. M. The route of enteric infection in normal mice. J Exp Med. 1974 May 1;139(5):1189–1203. doi: 10.1084/jem.139.5.1189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Coynault C., Robbe-Saule V., Norel F. Virulence and vaccine potential of Salmonella typhimurium mutants deficient in the expression of the RpoS (sigma S) regulon. Mol Microbiol. 1996 Oct;22(1):149–160. doi: 10.1111/j.1365-2958.1996.tb02664.x. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. 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]
  9. Foster J. W., Spector M. P. How Salmonella survive against the odds. Annu Rev Microbiol. 1995;49:145–174. doi: 10.1146/annurev.mi.49.100195.001045. [DOI] [PubMed] [Google Scholar]
  10. Foster J. W., Spector M. P. Phosphate starvation regulon of Salmonella typhimurium. J Bacteriol. 1986 May;166(2):666–669. doi: 10.1128/jb.166.2.666-669.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Galán J. E., Curtiss R., 3rd Cloning and molecular characterization of genes whose products allow Salmonella typhimurium to penetrate tissue culture cells. Proc Natl Acad Sci U S A. 1989 Aug;86(16):6383–6387. doi: 10.1073/pnas.86.16.6383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Galán J. E., Curtiss R., 3rd Expression of Salmonella typhimurium genes required for invasion is regulated by changes in DNA supercoiling. Infect Immun. 1990 Jun;58(6):1879–1885. doi: 10.1128/iai.58.6.1879-1885.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gentry D. R., Hernandez V. J., Nguyen L. H., Jensen D. B., Cashel M. Synthesis of the stationary-phase sigma factor sigma s is positively regulated by ppGpp. J Bacteriol. 1993 Dec;175(24):7982–7989. doi: 10.1128/jb.175.24.7982-7989.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Giannella R. A., Washington O., Gemski P., Formal S. B. Invasion of HeLa cells by Salmonella typhimurium: a model for study of invasiveness of Salmonella. J Infect Dis. 1973 Jul;128(1):69–75. doi: 10.1093/infdis/128.1.69. [DOI] [PubMed] [Google Scholar]
  15. Gulig P. A., Chiodo V. A. Genetic and DNA sequence analysis of the Salmonella typhimurium virulence plasmid gene encoding the 28,000-molecular-weight protein. Infect Immun. 1990 Aug;58(8):2651–2658. doi: 10.1128/iai.58.8.2651-2658.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Gulig P. A., Curtiss R., 3rd Cloning and transposon insertion mutagenesis of virulence genes of the 100-kilobase plasmid of Salmonella typhimurium. Infect Immun. 1988 Dec;56(12):3262–3271. doi: 10.1128/iai.56.12.3262-3271.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gulig P. A., Curtiss R., 3rd Plasmid-associated virulence of Salmonella typhimurium. Infect Immun. 1987 Dec;55(12):2891–2901. doi: 10.1128/iai.55.12.2891-2901.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Gulig P. A. Virulence plasmids of Salmonella typhimurium and other salmonellae. Microb Pathog. 1990 Jan;8(1):3–11. doi: 10.1016/0882-4010(90)90003-9. [DOI] [PubMed] [Google Scholar]
  19. HENLE G., DEINHARDT F. The establishment of strains of human cells in tissue culture. J Immunol. 1957 Jul;79(1):54–59. [PubMed] [Google Scholar]
  20. Hackett J., Kotlarski I., Mathan V., Francki K., Rowley D. The colonization of Peyer's patches by a strain of Salmonella typhimurium cured of the cryptic plasmid. J Infect Dis. 1986 Jun;153(6):1119–1125. doi: 10.1093/infdis/153.6.1119. [DOI] [PubMed] [Google Scholar]
  21. Hitchcock P. J., Brown T. M. Morphological heterogeneity among Salmonella lipopolysaccharide chemotypes in silver-stained polyacrylamide gels. J Bacteriol. 1983 Apr;154(1):269–277. doi: 10.1128/jb.154.1.269-277.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hoiseth S. K., Stocker B. A. Aromatic-dependent Salmonella typhimurium are non-virulent and effective as live vaccines. Nature. 1981 May 21;291(5812):238–239. doi: 10.1038/291238a0. [DOI] [PubMed] [Google Scholar]
  23. Iriarte M., Stainier I., Cornelis G. R. The rpoS gene from Yersinia enterocolitica and its influence on expression of virulence factors. Infect Immun. 1995 May;63(5):1840–1847. doi: 10.1128/iai.63.5.1840-1847.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. Kawahara K., Tsuchimoto M., Sudo K., Terakado N., Danbara H. Identification and mapping of mba regions of the Salmonella choleraesuis virulence plasmid pKDSC50 responsible for mouse bacteremia. Microb Pathog. 1990 Jan;8(1):13–21. doi: 10.1016/0882-4010(90)90004-a. [DOI] [PubMed] [Google Scholar]
  26. Kjelleberg S., Hermansson M., Mårdén P., Jones G. W. The transient phase between growth and nongrowth of heterotrophic bacteria, with emphasis on the marine environment. Annu Rev Microbiol. 1987;41:25–49. doi: 10.1146/annurev.mi.41.100187.000325. [DOI] [PubMed] [Google Scholar]
  27. Kowarz L., Coynault C., Robbe-Saule V., Norel F. The Salmonella typhimurium katF (rpoS) gene: cloning, nucleotide sequence, and regulation of spvR and spvABCD virulence plasmid genes. J Bacteriol. 1994 Nov;176(22):6852–6860. doi: 10.1128/jb.176.22.6852-6860.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Krause M., Roudier C., Fierer J., Harwood J., Guiney D. Molecular analysis of the virulence locus of the Salmonella dublin plasmid pSDL2. Mol Microbiol. 1991 Feb;5(2):307–316. doi: 10.1111/j.1365-2958.1991.tb02111.x. [DOI] [PubMed] [Google Scholar]
  29. LENNOX E. S. Transduction of linked genetic characters of the host by bacteriophage P1. Virology. 1955 Jul;1(2):190–206. doi: 10.1016/0042-6822(55)90016-7. [DOI] [PubMed] [Google Scholar]
  30. Lange R., Hengge-Aronis R. Growth phase-regulated expression of bolA and morphology of stationary-phase Escherichia coli cells are controlled by the novel sigma factor sigma S. J Bacteriol. 1991 Jul;173(14):4474–4481. doi: 10.1128/jb.173.14.4474-4481.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Lax A. J., Pullinger G. D., Baird G. D., Williamson C. M. The virulence plasmid of Salmonella dublin: detailed restriction map and analysis by transposon mutagenesis. J Gen Microbiol. 1990 Jun;136(6):1117–1123. doi: 10.1099/00221287-136-6-1117. [DOI] [PubMed] [Google Scholar]
  32. Lee C. A., Falkow S. The ability of Salmonella to enter mammalian cells is affected by bacterial growth state. Proc Natl Acad Sci U S A. 1990 Jun;87(11):4304–4308. doi: 10.1073/pnas.87.11.4304. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. 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]
  34. MOORE A. E., SABACHEWSKY L., TOOLAN H. W. Culture characteristics of four permanent lines of human cancer cells. Cancer Res. 1955 Oct;15(9):598–602. [PubMed] [Google Scholar]
  35. McCann M. P., Kidwell J. P., Matin A. The putative sigma factor KatF has a central role in development of starvation-mediated general resistance in Escherichia coli. J Bacteriol. 1991 Jul;173(13):4188–4194. doi: 10.1128/jb.173.13.4188-4194.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Mulvey M. R., Loewen P. C. Nucleotide sequence of katF of Escherichia coli suggests KatF protein is a novel sigma transcription factor. Nucleic Acids Res. 1989 Dec 11;17(23):9979–9991. doi: 10.1093/nar/17.23.9979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. 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]
  38. Nguyen L. H., Jensen D. B., Thompson N. E., Gentry D. R., Burgess R. R. In vitro functional characterization of overproduced Escherichia coli katF/rpoS gene product. Biochemistry. 1993 Oct 19;32(41):11112–11117. doi: 10.1021/bi00092a021. [DOI] [PubMed] [Google Scholar]
  39. Norel F., Coynault C., Miras I., Hermant D., Popoff M. Y. Cloning and expression of plasmid DNA sequences involved in Salmonella serotype typhimurium virulence. Mol Microbiol. 1989 Jun;3(6):733–743. doi: 10.1111/j.1365-2958.1989.tb00222.x. [DOI] [PubMed] [Google Scholar]
  40. Norel F., Robbe-Saule V., Popoff M. Y., Coynault C. The putative sigma factor KatF (RpoS) is required for the transcription of the Salmonella typhimurium virulence gene spvB in Escherichia coli. FEMS Microbiol Lett. 1992 Dec 1;78(2-3):271–276. doi: 10.1016/0378-1097(92)90039-q. [DOI] [PubMed] [Google Scholar]
  41. O'Neal C. R., Gabriel W. M., Turk A. K., Libby S. J., Fang F. C., Spector M. P. RpoS is necessary for both the positive and negative regulation of starvation survival genes during phosphate, carbon, and nitrogen starvation in Salmonella typhimurium. J Bacteriol. 1994 Aug;176(15):4610–4616. doi: 10.1128/jb.176.15.4610-4616.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Pardon P., Popoff M. Y., Coynault C., Marly J., Miras I. Virulence-associated plasmids of Salmonella serotype Typhimurium in experimental murine infection. Ann Inst Pasteur Microbiol. 1986 Jul-Aug;137B(1):47–60. doi: 10.1016/s0769-2609(86)80093-x. [DOI] [PubMed] [Google Scholar]
  43. Ralph P., Nakoinz I. Phagocytosis and cytolysis by a macrophage tumour and its cloned cell line. Nature. 1975 Oct 2;257(5525):393–394. doi: 10.1038/257393a0. [DOI] [PubMed] [Google Scholar]
  44. Rhen M., Virtanen M., Mäkelä P. H. Localization by insertion mutagenesis of a virulence-associated region on the Salmonella typhimurium 96 kilobase pair plasmid. Microb Pathog. 1989 Feb;6(2):153–158. doi: 10.1016/0882-4010(89)90018-1. [DOI] [PubMed] [Google Scholar]
  45. Rozdzinski E., Tuomanen E. Interactions of bacteria with leukocyte integrins. Methods Enzymol. 1994;236:333–345. doi: 10.1016/0076-6879(94)36025-1. [DOI] [PubMed] [Google Scholar]
  46. Spector M. P., Aliabadi Z., Gonzalez T., Foster J. W. Global control in Salmonella typhimurium: two-dimensional electrophoretic analysis of starvation-, anaerobiosis-, and heat shock-inducible proteins. J Bacteriol. 1986 Oct;168(1):420–424. doi: 10.1128/jb.168.1.420-424.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Spector M. P., Cubitt C. L. Starvation-inducible loci of Salmonella typhimurium: regulation and roles in starvation-survival. Mol Microbiol. 1992 Jun;6(11):1467–1476. doi: 10.1111/j.1365-2958.1992.tb00867.x. [DOI] [PubMed] [Google Scholar]
  48. Spector M. P., Park Y. K., Tirgari S., Gonzalez T., Foster J. W. Identification and characterization of starvation-regulated genetic loci in Salmonella typhimurium by using Mu d-directed lacZ operon fusions. J Bacteriol. 1988 Jan;170(1):345–351. doi: 10.1128/jb.170.1.345-351.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Sternberg N. L., Maurer R. Bacteriophage-mediated generalized transduction in Escherichia coli and Salmonella typhimurium. Methods Enzymol. 1991;204:18–43. doi: 10.1016/0076-6879(91)04004-8. [DOI] [PubMed] [Google Scholar]
  50. Tinge S. A., Curtiss R., 3rd Conservation of Salmonella typhimurium virulence plasmid maintenance regions among Salmonella serovars as a basis for plasmid curing. Infect Immun. 1990 Sep;58(9):3084–3092. doi: 10.1128/iai.58.9.3084-3092.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Tsai C. M., Frasch C. E. A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal Biochem. 1982 Jan 1;119(1):115–119. doi: 10.1016/0003-2697(82)90673-x. [DOI] [PubMed] [Google Scholar]
  52. Williamson C. M., Baird G. D., Manning E. J. A common virulence region on plasmids from eleven serotypes of Salmonella. J Gen Microbiol. 1988 Apr;134(4):975–982. doi: 10.1099/00221287-134-4-975. [DOI] [PubMed] [Google Scholar]
  53. Williamson C. M., Pullinger G. D., Lax A. J. Identification of an essential virulence region on Salmonella plasmids. Microb Pathog. 1988 Dec;5(6):469–473. doi: 10.1016/0882-4010(88)90008-3. [DOI] [PubMed] [Google Scholar]
  54. Wilmes-Riesenberg M. R., Foster J. W., Curtiss R., 3rd An altered rpoS allele contributes to the avirulence of Salmonella typhimurium LT2. Infect Immun. 1997 Jan;65(1):203–210. doi: 10.1128/iai.65.1.203-210.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]

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