Skip to main content
Infection and Immunity logoLink to Infection and Immunity
. 1993 Feb;61(2):705–713. doi: 10.1128/iai.61.2.705-713.1993

Stress induction of the virulence proteins (SpvA, -B, and -C) from native plasmid pSDL2 of Salmonella dublin.

S E Valone 1, G K Chikami 1, V L Miller 1
PMCID: PMC302783  PMID: 8380798

Abstract

The virulence region of the wild-type plasmid pSDL2 contained in Salmonella dublin is highly conserved among plasmids from several nontyphoid Salmonella serotypes and is essential for the development of systemic infection in BALB/c mice. Polyclonal antibodies against three proteins (SpvA, -B, and -C) expressed from a 4.1-kb EcoRI subclone of the plasmid virulence region were generated. These antibodies were used to detect expression of the Spv proteins when S. dublin was grown in vitro under stress-inducing conditions, such as nutrient deprivation and increased temperature, that the bacteria may encounter during the course of infection within the host. Glucose starvation resulted in expression of all three proteins shortly after the lag phase. When the bacteria were grown to the late-log phase without glucose, heat shock strongly induced expression of SpvA but not SpvB or SpvC. The addition of 0.2% glucose to the medium resulted in loss of expression of the proteins until the late-log to stationary phase. Iron limitation or lowered pH induced expression of the proteins during exponential growth even in the presence of glucose. Insertion mutations into the positive regulator gene spvR upstream from spvABC and insertions into spvA and spvC resulted in loss of expression of SpvA, -B, and -C, suggesting a complex regulation of expression. These studies define a variety of environmental conditions that induce expression of the Spv virulence proteins from the wild-type plasmid pSDL2 in S. dublin in vitro.

Full text

PDF
705

Images in this article

Selected References

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

  1. Abou-Zeid C., Filley E., Steele J., Rook G. A. A simple new method for using antigens separated by polyacrylamide gel electrophoresis to stimulate lymphocytes in vitro after converting bands cut from Western blots into antigen-bearing particles. J Immunol Methods. 1987 Apr 2;98(1):5–10. doi: 10.1016/0022-1759(87)90429-7. [DOI] [PubMed] [Google Scholar]
  2. Blaser M. J., Feldman R. A. From the centers for disease control. Salmonella bacteremia: reports to the Centers for Disease Control, 1968-1979. J Infect Dis. 1981 May;143(5):743–746. doi: 10.1093/infdis/143.5.743. [DOI] [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. 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. Chikami G. K., Fierer J., Guiney D. G. Plasmid-mediated virulence in Salmonella dublin demonstrated by use of a Tn5-oriT construct. Infect Immun. 1985 Nov;50(2):420–424. doi: 10.1128/iai.50.2.420-424.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cotter P. A., Gunsalus R. P. Oxygen, nitrate, and molybdenum regulation of dmsABC gene expression in Escherichia coli. J Bacteriol. 1989 Jul;171(7):3817–3823. doi: 10.1128/jb.171.7.3817-3823.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Daifuku R., Chikami G. K. Tn1725 transposon mutagenesis of 9-18 delta 7, an EcoRI deletion derivative of Salmonella dublin lane plasmid pSDL2. Infect Immun. 1991 Dec;59(12):4720–4723. doi: 10.1128/iai.59.12.4720-4723.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Fields P. I., Groisman E. A., Heffron F. A Salmonella locus that controls resistance to microbicidal proteins from phagocytic cells. Science. 1989 Feb 24;243(4894 Pt 1):1059–1062. doi: 10.1126/science.2646710. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Foster J. W., Hall H. K. Effect of Salmonella typhimurium ferric uptake regulator (fur) mutations on iron- and pH-regulated protein synthesis. J Bacteriol. 1992 Jul;174(13):4317–4323. doi: 10.1128/jb.174.13.4317-4323.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fürste J. P., Pansegrau W., Frank R., Blöcker H., Scholz P., Bagdasarian M., Lanka E. Molecular cloning of the plasmid RP4 primase region in a multi-host-range tacP expression vector. Gene. 1986;48(1):119–131. doi: 10.1016/0378-1119(86)90358-6. [DOI] [PubMed] [Google Scholar]
  13. Guiney D. G., Hasegawa P., Davis C. E. Plasmid transfer from Escherichia coli to Bacteroides fragilis: differential expression of antibiotic resistance phenotypes. Proc Natl Acad Sci U S A. 1984 Nov;81(22):7203–7206. doi: 10.1073/pnas.81.22.7203. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  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. 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]
  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. Heffernan E. J., Harwood J., Fierer J., Guiney D. The Salmonella typhimurium virulence plasmid complement resistance gene rck is homologous to a family of virulence-related outer membrane protein genes, including pagC and ail. J Bacteriol. 1992 Jan;174(1):84–91. doi: 10.1128/jb.174.1.84-91.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Krause M., Fang F. C., Guiney D. G. Regulation of plasmid virulence gene expression in Salmonella dublin involves an unusual operon structure. J Bacteriol. 1992 Jul;174(13):4482–4489. doi: 10.1128/jb.174.13.4482-4489.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  25. Larsson A., Nilsson B. O. Immunization with nanogram quantities of nitrocellulose-bound antigen, electroblotted from sodium dodecyl sulphate-polyacrylamide gels. Scand J Immunol. 1988 Mar;27(3):305–309. doi: 10.1111/j.1365-3083.1988.tb02351.x. [DOI] [PubMed] [Google Scholar]
  26. Matsui H., Abe A., Kawahara K., Terakado N., Danbara H. Positive regulator for the expression of Mba protein of the virulence plasmid, pKDSC50, of Salmonella choleraesuis. Microb Pathog. 1991 Jun;10(6):459–464. doi: 10.1016/0882-4010(91)90111-m. [DOI] [PubMed] [Google Scholar]
  27. Matsui H., Kawahara K., Terakado N., Danbara H. Nucleotide sequence of a gene encoding a 29 kDa polypeptide in mba region of the virulence plasmid, pKDSC50, of Salmonella choleraesuis. Nucleic Acids Res. 1990 Feb 25;18(4):1055–1055. doi: 10.1093/nar/18.4.1055. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Matsui H., Kawahara K., Terakado N., Danbara H. Nucleotide sequences of genes encoding 32 kDa and 70 kDa polypeptides in mba region of the virulence plasmid, pKDSc50, of Salmonella choleraesuis. Nucleic Acids Res. 1990 Apr 25;18(8):2181–2182. doi: 10.1093/nar/18.8.2181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Michiels T., Wattiau P., Brasseur R., Ruysschaert J. M., Cornelis G. Secretion of Yop proteins by Yersiniae. Infect Immun. 1990 Sep;58(9):2840–2849. doi: 10.1128/iai.58.9.2840-2849.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Miller S. I., Kukral A. M., Mekalanos J. J. A two-component regulatory system (phoP phoQ) controls Salmonella typhimurium virulence. Proc Natl Acad Sci U S A. 1989 Jul;86(13):5054–5058. doi: 10.1073/pnas.86.13.5054. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. Norel F., Pisano M. R., Nicoli J., Popoff M. Y. A plasmid-borne virulence region (2.8 kb) from Salmonella typhimurium contains two open reading frames. Res Microbiol. 1989 Nov-Dec;140(9):627–630. doi: 10.1016/0923-2508(89)90194-0. [DOI] [PubMed] [Google Scholar]
  33. Norel F., Pisano M. R., Nicoli J., Popoff M. Y. Nucleotide sequence of the plasmid-borne virulence gene mkfA encoding a 28 kDa polypeptide from Salmonella typhimurium. Res Microbiol. 1989 Mar-Apr;140(3):263–265. doi: 10.1016/0923-2508(89)90081-8. [DOI] [PubMed] [Google Scholar]
  34. Norel F., Pisano M. R., Nicoli J., Popoff M. Y. Nucleotide sequence of the plasmid-borne virulence gene mkfB from Salmonella typhimurium. Res Microbiol. 1989 Sep;140(7):455–457. doi: 10.1016/0923-2508(89)90066-1. [DOI] [PubMed] [Google Scholar]
  35. Pepe J. C., Miller V. L. The Yersinia enterocolitica inv gene product is an outer membrane protein that shares epitopes with Yersinia pseudotuberculosis invasin. J Bacteriol. 1990 Jul;172(7):3780–3789. doi: 10.1128/jb.172.7.3780-3789.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Pullinger G. D., Baird G. D., Williamson C. M., Lax A. J. Nucleotide sequence of a plasmid gene involved in the virulence of salmonellas. Nucleic Acids Res. 1989 Oct 11;17(19):7983–7983. doi: 10.1093/nar/17.19.7983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Roudier C., Fierer J., Guiney D. G. Characterization of translation termination mutations in the spv operon of the Salmonella virulence plasmid pSDL2. J Bacteriol. 1992 Oct;174(20):6418–6423. doi: 10.1128/jb.174.20.6418-6423.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Roudier C., Krause M., Fierer J., Guiney D. G. Correlation between the presence of sequences homologous to the vir region of Salmonella dublin plasmid pSDL2 and the virulence of twenty-two Salmonella serotypes in mice. Infect Immun. 1990 May;58(5):1180–1185. doi: 10.1128/iai.58.5.1180-1185.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Schellhorn H. E., Stones V. L. Regulation of katF and katE in Escherichia coli K-12 by weak acids. J Bacteriol. 1992 Jul;174(14):4769–4776. doi: 10.1128/jb.174.14.4769-4776.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Taira S., Baumann M., Riikonen P., Sukupolvi S., Rhen M. Amino-terminal sequence analysis of four plasmid-encoded virulence-associated proteins of Salmonella typhimurium. FEMS Microbiol Lett. 1991 Jan 15;61(2-3):319–323. doi: 10.1016/0378-1097(91)90573-s. [DOI] [PubMed] [Google Scholar]
  41. Taira S., Rhen M. Identification and genetic analysis of mkaA--a gene of the Salmonella typhimurium virulence plasmid necessary for intracellular growth. Microb Pathog. 1989 Sep;7(3):165–173. doi: 10.1016/0882-4010(89)90052-1. [DOI] [PubMed] [Google Scholar]
  42. Taira S., Rhen M. Molecular organization of genes constituting the virulence determinant on the Salmonella typhimurium 96 kilobase pair plasmid. FEBS Lett. 1989 Nov 6;257(2):274–278. doi: 10.1016/0014-5793(89)81551-0. [DOI] [PubMed] [Google Scholar]
  43. Taira S., Rhen M. Nucleotide sequence of mkaD, a virulence-associated gene of Salmonella typhimurium containing variable and constant regions. Gene. 1990 Sep 1;93(1):147–150. doi: 10.1016/0378-1119(90)90150-p. [DOI] [PubMed] [Google Scholar]
  44. Taira S., Riikonen P., Saarilahti H., Sukupolvi S., Rhen M. The mkaC virulence gene of the Salmonella serovar typhimurium 96 kb plasmid encodes a transcriptional activator. Mol Gen Genet. 1991 Sep;228(3):381–384. doi: 10.1007/BF00260630. [DOI] [PubMed] [Google Scholar]
  45. Valone S. E., Chikami G. K. Characterization of three proteins expressed from the virulence region of plasmid pSDL2 in Salmonella dublin. Infect Immun. 1991 Oct;59(10):3511–3517. doi: 10.1128/iai.59.10.3511-3517.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Vandenbosch J. L., Rabert D. K., Kurlandsky D. R., Jones G. W. Sequence analysis of rsk, a portion of the 95-kilobase plasmid of Salmonella typhimurium associated with resistance to the bactericidal activity of serum. Infect Immun. 1989 Mar;57(3):850–857. doi: 10.1128/iai.57.3.850-857.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Werner S. B., Humphrey G. L., Kamei I. Association between raw milk and human Salmonella dublin infection. Br Med J. 1979 Jul 28;2(6184):238–241. doi: 10.1136/bmj.2.6184.238. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. 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]
  49. Wray C., Sojka W. J. Reviews of the progress of dairy science: bovine salmonellosis. J Dairy Res. 1977 Jun;44(2):383–425. [PubMed] [Google Scholar]

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

RESOURCES