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. 1997 Jan;65(1):203–210. doi: 10.1128/iai.65.1.203-210.1997

An altered rpoS allele contributes to the avirulence of Salmonella typhimurium LT2.

M R Wilmes-Riesenberg 1, J W Foster 1, R Curtiss 3rd 1
PMCID: PMC174577  PMID: 8975913

Abstract

Virulent Salmonella typhimurium strains differ from the attenuated laboratory strain LT2 at the rpoS locus. It was previously shown that the rpoS gene in strain LT2 contains a rare UUG start codon (I. S. Lee, J. Lin, H. K. Hall, B. Bearson, and J. W. Foster, Mol. Microbiol. 17:155-167, 1995). This difference is responsible for the inability of LT2 to display a sustained log-phase acid tolerance response. We show that the altered rpoS allele (rpoS(LT2)) also affects the stationary-phase acid tolerance response in Salmonella. By transducing the rpoS(LT2) allele into virulent strain backgrounds and crossing wild-type rpoS allele into strain LT2, we demonstrate that the rpoS(LT2) allele contributes to the attenuation of strain LT2. We examined the effect of the rpoS allele on invasion and found that the rpoS status of the cell had no effect on the ability of the strains to invade intestinal epithelial cells in tissue culture. Enumeration of bacteria from tissues of infected mice indicated that the presence of the rpoS(LT2) allele affected the ability of S. typhimurium to reach the liver and spleen and to persist in several tissues at 6 days postinfection. This is likely due, at least in part, to a decrease in spv gene expression in these mutants. We demonstrate that strains containing the rpoS(LT2) allele are not only sensitive to pH 3.0 (acid stress) but are also sensitive to the DNA-damaging agent methyl methanesulfonate. However, these strains appear to survive stationary-phase and oxidative stresses as well as strains containing a wild-type rpoS allele. Despite an increased sensitivity to acid stress and DNA damage, strains containing either an rpoS-null mutation or the rpoS(LT2) allele survived in J774 cells and bone marrow-derived macrophages as well as did otherwise isogenic strains with a wild-type rpoS allele.

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Selected References

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  1. Abe A., Matsui H., Danbara H., Tanaka K., Takahashi H., Kawahara K. Regulation of spvR gene expression of Salmonella virulence plasmid pKDSC50 in Salmonella choleraesuis serovar Choleraesuis. Mol Microbiol. 1994 Jun;12(5):779–787. doi: 10.1111/j.1365-2958.1994.tb01064.x. [DOI] [PubMed] [Google Scholar]
  2. Abshire K. Z., Neidhardt F. C. Growth rate paradox of Salmonella typhimurium within host macrophages. J Bacteriol. 1993 Jun;175(12):3744–3748. doi: 10.1128/jb.175.12.3744-3748.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Benjamin W. H., Jr, Posey B. S., Briles D. E. Effects of in vitro growth phase on the pathogenesis of Salmonella typhimurium in mice. J Gen Microbiol. 1986 May;132(5):1283–1295. doi: 10.1099/00221287-132-5-1283. [DOI] [PubMed] [Google Scholar]
  4. Benjamin W. H., Jr, Yother J., Hall P., Briles D. E. The Salmonella typhimurium locus mviA regulates virulence in Itys but not Ityr mice: functional mviA results in avirulence; mutant (nonfunctional) mviA results in virulence. J Exp Med. 1991 Nov 1;174(5):1073–1083. doi: 10.1084/jem.174.5.1073. [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. 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]
  7. 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]
  8. Ernst R. K., Dombroski D. M., Merrick J. M. Anaerobiosis, type 1 fimbriae, and growth phase are factors that affect invasion of HEp-2 cells by Salmonella typhimurium. Infect Immun. 1990 Jun;58(6):2014–2016. doi: 10.1128/iai.58.6.2014-2016.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. 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]
  11. Foster J. W., Hall H. K. Adaptive acidification tolerance response of Salmonella typhimurium. J Bacteriol. 1990 Feb;172(2):771–778. doi: 10.1128/jb.172.2.771-778.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Galán J. E., Curtiss R., 3rd Virulence and vaccine potential of phoP mutants of Salmonella typhimurium. Microb Pathog. 1989 Jun;6(6):433–443. doi: 10.1016/0882-4010(89)90085-5. [DOI] [PubMed] [Google Scholar]
  14. Garcia-del Portillo F., Foster J. W., Finlay B. B. Role of acid tolerance response genes in Salmonella typhimurium virulence. Infect Immun. 1993 Oct;61(10):4489–4492. doi: 10.1128/iai.61.10.4489-4492.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gulig P. A., Caldwell A. L., Chiodo V. A. Identification, genetic analysis and DNA sequence of a 7.8-kb virulence region of the Salmonella typhimurium virulence plasmid. Mol Microbiol. 1992 May;6(10):1395–1411. doi: 10.1111/j.1365-2958.1992.tb00860.x. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Gulig P. A., Danbara H., Guiney D. G., Lax A. J., Norel F., Rhen M. Molecular analysis of spv virulence genes of the Salmonella virulence plasmids. Mol Microbiol. 1993 Mar;7(6):825–830. doi: 10.1111/j.1365-2958.1993.tb01172.x. [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. 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. Heiskanen P., Taira S., Rhen M. Role of rpoS in the regulation of Salmonella plasmid virulence (spv) genes. FEMS Microbiol Lett. 1994 Oct 15;123(1-2):125–130. doi: 10.1111/j.1574-6968.1994.tb07211.x. [DOI] [PubMed] [Google Scholar]
  21. Ivanova A., Renshaw M., Guntaka R. V., Eisenstark A. DNA base sequence variability in katF (putative sigma factor) gene of Escherichia coli. Nucleic Acids Res. 1992 Oct 25;20(20):5479–5480. doi: 10.1093/nar/20.20.5479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kusters J. G., Mulders-Kremers G. A., van Doornik C. E., van der Zeijst B. A. Effects of multiplicity of infection, bacterial protein synthesis, and growth phase on adhesion to and invasion of human cell lines by Salmonella typhimurium. Infect Immun. 1993 Dec;61(12):5013–5020. doi: 10.1128/iai.61.12.5013-5020.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. LURIA S. E., BURROUS J. W. Hybridization between Escherichia coli and Shigella. J Bacteriol. 1957 Oct;74(4):461–476. doi: 10.1128/jb.74.4.461-476.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. Lee I. S., Lin J., Hall H. K., Bearson B., Foster J. W. The stationary-phase sigma factor sigma S (RpoS) is required for a sustained acid tolerance response in virulent Salmonella typhimurium. Mol Microbiol. 1995 Jul;17(1):155–167. doi: 10.1111/j.1365-2958.1995.mmi_17010155.x. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Lonetto M., Gribskov M., Gross C. A. The sigma 70 family: sequence conservation and evolutionary relationships. J Bacteriol. 1992 Jun;174(12):3843–3849. doi: 10.1128/jb.174.12.3843-3849.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. 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]
  31. 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]
  32. Riesenberg-Wilmes M. R., Bearson B., Foster J. W., Curtis R., 3rd Role of the acid tolerance response in virulence of Salmonella typhimurium. Infect Immun. 1996 Apr;64(4):1085–1092. doi: 10.1128/iai.64.4.1085-1092.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Robbe-Saule V., Coynault C., Norel F. The live oral typhoid vaccine Ty21a is a rpoS mutant and is susceptible to various environmental stresses. FEMS Microbiol Lett. 1995 Feb 15;126(2):171–176. doi: 10.1111/j.1574-6968.1995.tb07412.x. [DOI] [PubMed] [Google Scholar]
  34. Schmieger H. Phage P22-mutants with increased or decreased transduction abilities. Mol Gen Genet. 1972;119(1):75–88. doi: 10.1007/BF00270447. [DOI] [PubMed] [Google Scholar]
  35. Tanaka K., Takayanagi Y., Fujita N., Ishihama A., Takahashi H. Heterogeneity of the principal sigma factor in Escherichia coli: the rpoS gene product, sigma 38, is a second principal sigma factor of RNA polymerase in stationary-phase Escherichia coli. Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3511–3515. doi: 10.1073/pnas.90.8.3511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. VOGEL H. J., BONNER D. M. Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956 Jan;218(1):97–106. [PubMed] [Google Scholar]
  37. Valone S. E., Chikami G. K., Miller V. L. Stress induction of the virulence proteins (SpvA, -B, and -C) from native plasmid pSDL2 of Salmonella dublin. Infect Immun. 1993 Feb;61(2):705–713. doi: 10.1128/iai.61.2.705-713.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Vladoianu I. R., Chang H. R., Pechère J. C. Expression of host resistance to Salmonella typhi and Salmonella typhimurium: bacterial survival within macrophages of murine and human origin. Microb Pathog. 1990 Feb;8(2):83–90. doi: 10.1016/0882-4010(90)90072-x. [DOI] [PubMed] [Google Scholar]
  39. 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]

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