1887

Abstract

The genes of and other non-typhoidal serovars are essential for efficient systemic infection beyond the intestines in orally inoculated mice as a model for enteric fever. These virulence genes are not significantly expressed by salmonellae during exponential growth in broth but are induced when the bacteria enter the stationary phase of growth. Using RNase protection analysis to directly measure mRNA from the virulence plasmid of we found that was maximally induced in an SpvR- and RpoS-dependent manner during exponential growth in Intracellular Salts Medium, which mimics the intracellular environment of mammalian cells. A cloned operon fusion in was induced intracellularly in peritoneal cells of mice, correlating intracellular gene expression with intracellular function of the genes in infected mice. was also induced intracellularly within both Henle-407 intestinal epithelial cells and J774.A1 macrophage-like cells when the bacteria were replicating with exponential kinetics. Prevention of invasion of salmonellae with cytochalasin inhibited induction within tissue culture cells, indicating that salmonellae must be internalized for to be induced. The fusion was not induced by salmonellae in extracellular fluid of the peritoneal cavity or in serum. Since induction of the spv genes occurs intracellularly during exponential growth of salmonellae, cessation of growth may not be the most relevant inducing signal for spv gene expression.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-143-12-3827
1997-12-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/micro/143/12/mic-143-12-3827.html?itemId=/content/journal/micro/10.1099/00221287-143-12-3827&mimeType=html&fmt=ahah

References

  1. Abe A., Matsui H., Danbara H., Tanaka K., Takahashi H., Kawahara K. 1994; Regulation of spvR gene expression of Salmonella virulence plasmid pKDSC50 in Salmonella choleraesuis serovar choleraesuis.. Mol Microbiol 12:779–787
    [Google Scholar]
  2. Abshire K. Z., Neidhardt F. C. 1993; Growth rate paradox of Salmonella typhimurium within host macrophages.. J Bacteriol 175:3744–3748
    [Google Scholar]
  3. Ausubel F.M., Brent R., Kingston R.E., Moore D.D., Seidman J.G., Smith J.A., Struhl K. 1993 Current Protocols in Molecular Biology. New Yor: Wiley Interscience;
    [Google Scholar]
  4. Bohannon D.E., Connell N., Keener J., Tormo A., Espinosa-Urgel M., Mercedes Zambrano M., Kolter R. 1991; Stationary-phase-inducible ‘gearbox’ promoters: differential effects of katF mutations and role of σ70.. J Bacteriol 173:4482–4492
    [Google Scholar]
  5. Caldwell A.L., Gulig P.A. 1991; The Salmonella typhimurium virulence plasmid encodes a positive regulator of a plasmid-encoded virulence gene.. J Bacteriol 173:7176–7185
    [Google Scholar]
  6. Chen C.Y., Buchmeier N.A., Libby S., Fang F.C., Krause M., Guiney D.G. 1995; Central regulatory role for the RpoS sigma factor in expression of Salmonella dublin plasmid virulence genes.. J Bacteriol 111:5303–5309
    [Google Scholar]
  7. Chen C.Y., Eckmann L., Libby S.J., Fang F.C., Okamoto S., Kagnoff M.F., Fierer J., Guiney D.G. 1996a; Expression of Salmonella typhimurium rpoS and rpoS-dependent genes in the intracellular environment of eukarytotic cells.. Infect Immun 64:4739–4743
    [Google Scholar]
  8. Chen L.M., Kaniga K., Galan J.E. 1996b; Salmonella spp. are cytotoxic for cultured macrophages.. Mol Microbiol 21:1101–1115
    [Google Scholar]
  9. Coynault C., Robbe-Saule V., Popoff M.Y., Norel F. 1992; Growth phase and SpvR regulation of transcription of Salmonella typhimurium spvABC virulence genes.. Microb Pathog 13:133–143
    [Google Scholar]
  10. Deretic V., Gill J.F., Chakrabarty A.M. 1987; Gene algD coding for GDP mannose dehydrogenase is transcriptionally activated in mucoid Pseudomonas aeruginosa.. J Bacteriol 169:351–358
    [Google Scholar]
  11. Donnenberg M.S., Kaper J.B. 1991; Construction of an eae deletion mutant of enteropathogenic Escherichia coli by using a positive-selection suicide vector.. Infect Immun 59:4310–4317
    [Google Scholar]
  12. El-Gedaily A., Paesold G., Chen C.-Y., Guiney D.G., Krause M. 1997; Plasmid virulence gene expression induced by short-chain fatty acids in Salmonella dublin: Identification of rpoS-dependent and rpoS-independent mechanisms.. J Bacteriol 179:1409–1412
    [Google Scholar]
  13. Errington J. 1986; A general method for fusion of the Escherichia coli lacZ gene to chromosomal genes in Bacillus subtilis.. J Gen Microbiol 132:2953–2966
    [Google Scholar]
  14. Fang F.C., Krause M., Roudier C., Fierer J., Guiney D.G. 1991; Growth regulation of a Salmonella plasmid gene essential for virulence.. J Bacteriol 173:6783–6789
    [Google Scholar]
  15. Fang F.C., Libby S.J., Buchmeier N.A., Loewen P.C., Switala J., Harwood J., Guiney D.G. 1992; The alternative sigma factor KatF (RpoS) regulates Salmonella virulence.. Proc Natl Acad Sci USA 8911978–11982
    [Google Scholar]
  16. Fierer J., Krause M., Tauxe R., Guiney D. 1992; Salmonella typhimurium bacteremia: association with the virulence plasmid.. J Infect Dis 166:639–642
    [Google Scholar]
  17. Fierer J., Eckmann L., Fang F., Pfeifer C., Finlay B.B., Guiney D. 1993; Expression of the Salmonella virulence plasmid gene spvB in cultured macrophages and nonphagocytic cells.. Infect Immun 61:5231–5236
    [Google Scholar]
  18. Finlay B.B., Falkow S. 1988; Comparison of the invasion strategies used by Salmonella cholerae-suis, Shigella flexneri, and Yersinia enterocolitica to enter cultured animal cells: endosome acidification is not required for bacterial invasion or intracellular replication.. Biochimie 70:1089–1099
    [Google Scholar]
  19. Finlay B.B., Falkow S. 1989; Common themes in microbial pathogenesis.. Microbiol Rev 53:210–230
    [Google Scholar]
  20. Fleming T.J., Fleming M.L., Malek T.R. 1993; Selective expression of Ly-6G on myeloid lineage cells in mouse bone marrow. RB6-8C5 mAb to granulocyte-differentiation antigen (Gr-1) detects members of the Ly-6 family.. J Immunol 151:2399–2408
    [Google Scholar]
  21. Groat R.G., Schultz J.E., Zychlinsky E., Bockman A., Matin A. 1986; Starvation proteins in Escherichia coli: kinetics of synthesis and role in starvation survival.. J Bacteriol 168:486–493
    [Google Scholar]
  22. Guiney D.G., Libby S., Fang F.C., Krause M., Fierer J. 1995; Growth-phase regulation of plasmid virulence genes in Salmonella.. Trends Microbiol 3:275–279
    [Google Scholar]
  23. Gulig P.A., Curtiss R. III 1987; Plasmid-associated virulence of Salmonella typhimurium.. Infect Immun 55:2891–2901
    [Google Scholar]
  24. Gulig P.A., Doyle T.J. 1993; The Salmonella typhimurium virulence plasmid increases the growth rate of salmonellae in mice.. Infect Immun 61:504–511
    [Google Scholar]
  25. Gulig P.A., Caldwell A.L., Chiodo V.A. 1992; Identification, genetic analysis, and DNA sequence of a 7·8-kilobase virulence region of the Salmonella typhimurium virulence plasmid.. Mol Microbiol 6:1395–1411
    [Google Scholar]
  26. Gulig P.A., Danbara H., Guiney D.G., Lax A.J., Norel F., Rhen M. 1993; Molecular analysis of virulence genes of the salmonella virulence plasmids.. Mol Microbiol 7:825–830
    [Google Scholar]
  27. Gutierrez C., Barondess J., Manoil C., Beckwith J. 1987; The use of transposon Tnp^oA to detect genes for cell envelope proteins subject to a common regulatory stimulus. Analysis of osmotically regulated genes in Escherichia coli.. J Mol Biol 195:289–297
    [Google Scholar]
  28. Headley V.L., Payne S.M. 1990; Differential protein expression by Shigella flexneri in intracellular and extracellular environments.. Proc Natl Acad Sci USA 874179–4183
    [Google Scholar]
  29. Heiskanen P., Taira S., Rhen M. 1994; Role of rpoS in the regulation of Salmonella plasmid virulence (spv) genes.. FEMS Microbiol Lett 123:125–130
    [Google Scholar]
  30. Heithoff D.M., Conner C.P., Hanna P.C., Julio S.M., Hentschel U., Mahan M.J. 1997; Bacterial infection as assessed by in vivo gene expression.. Proc Natl Acad Sci USA 94934–939
    [Google Scholar]
  31. Hengge-Aronis R. 1996; Back to log phase: σs as a global regulator in the osmotic control of gene expression in Escherichia coli.. Mol Microbiol 21:887–893
    [Google Scholar]
  32. Hengge-Aronis R., Klein W., Lange R., Rimmele M., Boos W. 1991; Trehalose synthesis genes are controlled by the putative sigma factor encoded by rpoS and are involved in stationary- phase thermotolerance in Escherichia coli.. J Bacteriol 173:7918–7924
    [Google Scholar]
  33. Jenkins D.E., Schultz J.E., Matin A. 1988; Starvation-induced cross-protection against heat or H2O2 challenge in Escherichia coli.. J Bacteriol 170:3910–3914
    [Google Scholar]
  34. Jishage M., Iwata A., Ueda S., Ishihama A. 1996; Regulation of RNA polymerase sigma subunit synthesis in Escherichia coli: intracellular levels of four species of sigma subunit under various growth conditions.. J Bacteriol 178:5447–5451
    [Google Scholar]
  35. Kowarz L., Coynault C., Robbe Saule V., Norel F. 1994; The Salmonella typhimurium katF (rpoS) gene: cloning, nucleotide sequence, and regulation of spvR and spvABCD virulence plasmid genes.. J Bacteriol 176:6852–6860
    [Google Scholar]
  36. Krause M., Fang F.C., Guiney D.G. 1992; Regulation of plasmid virulence gene expression in Salmonella dublin involves an unusual operon structure.. J Bacteriol 174:4482–4489
    [Google Scholar]
  37. Lange R., Hengge-Aronis R. 1991; 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 173:4474–4481
    [Google Scholar]
  38. Lennox E.S. 1955; Transduction of linked genetic characters of the host by bacteriophage PI.. Virology 1:190–206
    [Google Scholar]
  39. McCann M.P., Kidwell J.P., Matin A. 1991; The putative sigma factor KatF has a central role in development of starvation-mediated general resistance in Escherichia coli.. J Bacteriol 173:4188–4194
    [Google Scholar]
  40. McKnight A.J., Macfarlane A.J., Dri P., Turley L., Willis A.C., Gordon S. 1996; Molecular cloning of F4/80, a murine macrophage-restricted cell surface glycoprotein with homology to the G-protein-linked transmembrane 7 hormone receptor family.. J Biol Chem 271:486–489
    [Google Scholar]
  41. Maniatis T., Fritsch E.F., Sambrook J. 1982 Molecular Cloning: a Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  42. Matsui H., Abe A., Kawahara K., Terakado N., Danbara H. 1991; Positive regulator for the expression of Mba protein of the virulence plasmid, pKDSC50, of Salmonella cboleraesuis.. Microb Pathog 10:459–464
    [Google Scholar]
  43. Matsui H., Abe A., Suzuki S., Kijima M., Tamura Y., Nakamura M., Kawahara K., Danbara H. 1993; Molecular mechanism of the regulation of expression of plasmid-encoded mouse bacteremia (mba) genes in Salmonella serovar choleraesuis.. Mol Gen Genet 236:219–226
    [Google Scholar]
  44. Miller J.H. 1972 Experiments in Molecular Genetics. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  45. Monack D.M., Raupach B., Hromockyj A.E., Falkow S. 1996; Salmonella typhimurium invasion induces apoptosis in infected macrophages.. Proc Natl Acad Sci USA 939833–9838
    [Google Scholar]
  46. Montenegro M.A., Morelli G., Helmuth R. 1991; Heteroduplex analysis of Salmonella virulence plasmids and their prevalence in isolates of defined sources.. Microb Pathog 11:391–397
    [Google Scholar]
  47. Muffler A., Barth M., Marchall C., Hengge-Aronis R. 1997; Heat shock regulation of σs turnover: a role for DnaK and relationship between stress responses mediated by σs and σ32 in Escherichia coli.. J Bacterial 179:445–452
    [Google Scholar]
  48. Norel F., Robbe Saule V., Popoff M.Y., Coynault C. 1992; The putative sigma factor KatF (RpoS) is required for the transcription of the Salmonella typhimurium virulence gene spvB in Escherichia coli.. FEMS Microbiol Lett 78:271–276
    [Google Scholar]
  49. O´Byrne C.P., Dorman C.J. 1994a; The spv virulence operon of Salmonella typhimurium LT2 is regulated negatively by the cyclic AMP (cAMP)-cAMP receptor protein system.. J Bacteriol 176:905–912
    [Google Scholar]
  50. O´Byrne C.P., Dorman C.J. 1994b; Transcription of the Salmonella typhimurium spv virulence locus is regulated negatively by the nucleoid-associated protein H-NS.. FEMS Microbiol Lett 121:99–105
    [Google Scholar]
  51. O´Neal C.R., Gabriel W.M., Turk A.K., Libby S.J., Fang F.C., Spector M.P. 1994; 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 176:4610–4616
    [Google Scholar]
  52. Pollack C., Straley S.C., Klempner M.S. 1986; Probing the phagolysosomal environment of human macrophages with a Ca2+-responsive operon fusion in Yersinia pestis.. Nature 322:834–836
    [Google Scholar]
  53. Rhen M., Riikonen P., Taira S. 1993; Transcriptional regulation of Salmonella enterica virulence plasmid genes in cultured macrophages.. Mol Microbiol 10:45–56
    [Google Scholar]
  54. Riikonen P., Makela P.H., Saarilahti H., Sukupolvi S., Taira S., Rhen M. 1992; The virulence plasmid does not contribute to growth of Salmonella in cultured murine macrophages.. Microb Pathog 13:281–291
    [Google Scholar]
  55. Rogers J.A., Matsui H., Gulig P.A. 1995; spv A of the Salmonella typhimurium virulence plasmid can act as a transcriptional repressor of spvR and is not essential for virulence in orally inoculated mice.. Abstr 95th Annu Meet Am Soc Microbiol B304.
    [Google Scholar]
  56. Schmeiger H. 1972; Phage P22-mutants with increased or decreased transduction abilities.. Mol Gen Genet 119:75–88
    [Google Scholar]
  57. Spink J.M., Pullinger G.D., Wood M.W., Lax A.J. 1994; Regulation of spvR, the positive regulatory gene of Salmonella plasmid virulence genes.. FEMS Microbiol Lett 116:113–121
    [Google Scholar]
  58. Taira S., Riikonen P., Saarilahti H., Sukupolvi S., Rhen M. 1991; The mkaC virulence gene of the Salmonella serovar typhimurium 96 kb plasmid encodes a transcriptional activator.. Mol Gen Genet 228:381–384
    [Google Scholar]
  59. Taira S., Heiskanen P., Hurme R., Heikkila H., Riikonen P., Rhen M. 1995; Evidence for functional polymorphism of the spvR gene regulating virulence gene expression in Salmonella.. Mol Gen Genet 246:437–444
    [Google Scholar]
  60. Valone S.E., Chikami G.K., Miller V.L. 1993; Stress induction of the virulence proteins (SpvA, -B, and -C) from native plasmid pSDL2 of Salmonella dublin.. Infect Immun 61:705–713
    [Google Scholar]
  61. Wilkins E.G.L., Roberts C. 1988; Extraintestinal salmonellosis.. Epidemiol Infect 100:361–368
    [Google Scholar]
  62. Yanisch-Perron C., Vieira J., Messing J. 1985; Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mpl8 and pUC19 vectors.. Gene 33:103–119
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-143-12-3827
Loading
/content/journal/micro/10.1099/00221287-143-12-3827
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error