Abstract
It is established that the medically significant yersiniae require the presence of physiological levels of Ca2+ (ca. 2.5 mM) for sustained growth at 37 degrees C and that this nutritional requirement is mediated by a shared ca. 70-kb Lcr plasmid. The latter also encodes virulence factors (Yersinia outer membrane proteins [Yops] and V antigen) known to be selectively synthesized in vitro at 37 degrees C in Ca(2+)-deficient medium. In this study, cells of Yersinia pestis KIM were first starved for Ca2+ at 37 degrees C to prevent synthesis of bulk vegetative protein and then, after cell division had ceased, pulsed with [35S]methionine. After sufficient chase to ensure plasminogen activator-mediated degradation of Yops, the remaining major radioactive peptides were separated by conventional chromatographic methods and identified as Lcr plasmid-encoded V antigen and LcrH (and possibly LcrG), ca. 10-kb Pst plasmid-encoded pesticin and plasminogen activator, ca. 100-kb Tox plasmid-encoded fraction 1 (capsular) antigen and murine exotoxin, and chromosomally encoded antigen 4 (pH 6 antigen) and antigen 5 (a novel hemin-rich peptide possessing modest catalase activity but not superoxide dismutase activity). Also produced at high concentration was a chromosome-encoded GroEL-like chaperone protein. Accordingly, the transcriptional block preventing synthesis of bulk vegetative protein at 37 degrees C in Ca(2+)-deficient medium may not apply to genes encoding virulence factors or to highly conserved GroEL (known in other species to utilize a secondary stress-induced sigma factor).
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- BACON G. A., BURROWS T. W. The basis of virulence in Pasteurella pestis: an antigen determining virulence. Br J Exp Pathol. 1956 Oct;37(5):481–493. [PMC free article] [PubMed] [Google Scholar]
- BAKER E. E., SOMMER H., FOSTER L. E., MEYER E., MEYER K. F. Studies on immunization against plague. I. The isolation and characterization of the soluble antigen of Pasteurella pestis. J Immunol. 1952 Feb;68(2):131–145. [PubMed] [Google Scholar]
- BEERS R. F., Jr, SIZER I. W. A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. J Biol Chem. 1952 Mar;195(1):133–140. [PubMed] [Google Scholar]
- BICHOWSKY-SLOMNICKI L., BEN-EFRAIM S. BIOLOGICAL ACTIVITIES IN EXTRACTS OF PASTEURELLA PESTIS AND THEIR RELATION TO THE "PH 6 ANTIGEN". J Bacteriol. 1963 Jul;86:101–111. doi: 10.1128/jb.86.1.101-111.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
- BRUBAKER R. R., SURGALLA M. J. THE EFFECT OF CA++ AND MG++ ON LYSIS, GROWTH, AND PRODUCTION OF VIRULENCE ANTIGENS BY PASTEURELLA PESTIS. J Infect Dis. 1964 Feb;114:13–25. doi: 10.1093/infdis/114.1.13. [DOI] [PubMed] [Google Scholar]
- Barve S. S., Straley S. C. lcrR, a low-Ca2(+)-response locus with dual Ca2(+)-dependent functions in Yersinia pestis. J Bacteriol. 1990 Aug;172(8):4661–4671. doi: 10.1128/jb.172.8.4661-4671.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bergman T., Håkansson S., Forsberg A., Norlander L., Macellaro A., Bäckman A., Bölin I., Wolf-Watz H. Analysis of the V antigen lcrGVH-yopBD operon of Yersinia pseudotuberculosis: evidence for a regulatory role of LcrH and LcrV. J Bacteriol. 1991 Mar;173(5):1607–1616. doi: 10.1128/jb.173.5.1607-1616.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brubaker R. R. Factors promoting acute and chronic diseases caused by yersiniae. Clin Microbiol Rev. 1991 Jul;4(3):309–324. doi: 10.1128/cmr.4.3.309. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brubaker R. R. Growth of Pasteurella pseudotuberculosis in simulated intracellular and extracellular environments. J Infect Dis. 1967 Dec;117(5):403–417. doi: 10.1093/infdis/117.5.403. [DOI] [PubMed] [Google Scholar]
- Brubaker R. R., Sample A. K., Yu D. Z., Zahorchak R. J., Hu P. C., Fowler J. M. Proteolysis of V antigen from Yersinia pestis. Microb Pathog. 1987 Jan;2(1):49–62. doi: 10.1016/0882-4010(87)90114-8. [DOI] [PubMed] [Google Scholar]
- Bölin I., Portnoy D. A., Wolf-Watz H. Expression of the temperature-inducible outer membrane proteins of yersiniae. Infect Immun. 1985 Apr;48(1):234–240. doi: 10.1128/iai.48.1.234-240.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- CRUMPTON M. J., DAVIES D. A. An antigenic analysis of Pasteurella pestis by diffusion of antigens and antibodies in agar. Proc R Soc Lond B Biol Sci. 1956 Mar 27;144(918):109–134. doi: 10.1098/rspb.1956.0021. [DOI] [PubMed] [Google Scholar]
- Charnetzky W. T., Brubaker R. R. RNA synthesis in Yersinia pestis during growth restriction in calcium-deficient medium. J Bacteriol. 1982 Mar;149(3):1089–1095. doi: 10.1128/jb.149.3.1089-1095.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Christman M. F., Morgan R. W., Jacobson F. S., Ames B. N. Positive control of a regulon for defenses against oxidative stress and some heat-shock proteins in Salmonella typhimurium. Cell. 1985 Jul;41(3):753–762. doi: 10.1016/s0092-8674(85)80056-8. [DOI] [PubMed] [Google Scholar]
- Cornelis G., Laroche Y., Balligand G., Sory M. P., Wauters G. Yersinia enterocolitica, a primary model for bacterial invasiveness. Rev Infect Dis. 1987 Jan-Feb;9(1):64–87. doi: 10.1093/clinids/9.1.64. [DOI] [PubMed] [Google Scholar]
- Ellis J. Proteins as molecular chaperones. 1987 Jul 30-Aug 5Nature. 328(6129):378–379. doi: 10.1038/328378a0. [DOI] [PubMed] [Google Scholar]
- Forsberg A., Bölin I., Norlander L., Wolf-Watz H. Molecular cloning and expression of calcium-regulated, plasmid-coded proteins of Y. pseudotuberculosis. Microb Pathog. 1987 Feb;2(2):123–137. doi: 10.1016/0882-4010(87)90104-5. [DOI] [PubMed] [Google Scholar]
- Gorbunoff M. J., Timasheff S. N. The interaction of proteins with hydroxyapatite. III. Mechanism. Anal Biochem. 1984 Feb;136(2):440–445. doi: 10.1016/0003-2697(84)90241-0. [DOI] [PubMed] [Google Scholar]
- Grossman A. D., Erickson J. W., Gross C. A. The htpR gene product of E. coli is a sigma factor for heat-shock promoters. Cell. 1984 Sep;38(2):383–390. doi: 10.1016/0092-8674(84)90493-8. [DOI] [PubMed] [Google Scholar]
- Grossman A. D., Taylor W. E., Burton Z. F., Burgess R. R., Gross C. A. Stringent response in Escherichia coli induces expression of heat shock proteins. J Mol Biol. 1985 Nov 20;186(2):357–365. doi: 10.1016/0022-2836(85)90110-x. [DOI] [PubMed] [Google Scholar]
- Heesemann J., Gross U., Schmidt N., Laufs R. Immunochemical analysis of plasmid-encoded proteins released by enteropathogenic Yersinia sp. grown in calcium-deficient media. Infect Immun. 1986 Nov;54(2):561–567. doi: 10.1128/iai.54.2.561-567.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hendrix R. W. Purification and properties of groE, a host protein involved in bacteriophage assembly. J Mol Biol. 1979 Apr 15;129(3):375–392. doi: 10.1016/0022-2836(79)90502-3. [DOI] [PubMed] [Google Scholar]
- Hightower L. E. Heat shock, stress proteins, chaperones, and proteotoxicity. Cell. 1991 Jul 26;66(2):191–197. doi: 10.1016/0092-8674(91)90611-2. [DOI] [PubMed] [Google Scholar]
- Hu P. C., Brubaker R. R. Characterization of pesticin. Separation of antibacterial activities. J Biol Chem. 1974 Aug 10;249(15):4749–4753. [PubMed] [Google Scholar]
- Hu P. C., Yang G. C., Brubaker R. R. Specificity, induction, and absorption of pesticin. J Bacteriol. 1972 Oct;112(1):212–219. doi: 10.1128/jb.112.1.212-219.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kumamoto C. A. Molecular chaperones and protein translocation across the Escherichia coli inner membrane. Mol Microbiol. 1991 Jan;5(1):19–22. doi: 10.1111/j.1365-2958.1991.tb01821.x. [DOI] [PubMed] [Google Scholar]
- LAWTON W. D., FUKUI G. M., SURGALLA M. J. Studies on the antigens of Pasteurella pestis and Pasteurella pseudotuberculosis. J Immunol. 1960 May;84:475–479. [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Lecker S., Lill R., Ziegelhoffer T., Georgopoulos C., Bassford P. J., Jr, Kumamoto C. A., Wickner W. Three pure chaperone proteins of Escherichia coli--SecB, trigger factor and GroEL--form soluble complexes with precursor proteins in vitro. EMBO J. 1989 Sep;8(9):2703–2709. doi: 10.1002/j.1460-2075.1989.tb08411.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lindler L. E., Klempner M. S., Straley S. C. Yersinia pestis pH 6 antigen: genetic, biochemical, and virulence characterization of a protein involved in the pathogenesis of bubonic plague. Infect Immun. 1990 Aug;58(8):2569–2577. doi: 10.1128/iai.58.8.2569-2577.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lucier T. S., Brubaker R. R. Determination of genome size, macrorestriction pattern polymorphism, and nonpigmentation-specific deletion in Yersinia pestis by pulsed-field gel electrophoresis. J Bacteriol. 1992 Apr;174(7):2078–2086. doi: 10.1128/jb.174.7.2078-2086.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Matin A. The molecular basis of carbon-starvation-induced general resistance in Escherichia coli. Mol Microbiol. 1991 Jan;5(1):3–10. doi: 10.1111/j.1365-2958.1991.tb01819.x. [DOI] [PubMed] [Google Scholar]
- Mehigh R. J., Sample A. K., Brubaker R. R. Expression of the low calcium response in Yersinia pestis. Microb Pathog. 1989 Mar;6(3):203–217. doi: 10.1016/0882-4010(89)90070-3. [DOI] [PubMed] [Google Scholar]
- Michiels T., Cornelis G. R. Secretion of hybrid proteins by the Yersinia Yop export system. J Bacteriol. 1991 Mar;173(5):1677–1685. doi: 10.1128/jb.173.5.1677-1685.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Montie T. C., Montie D. B. Protein toxins of Pasteurella pestis. Subunit composition and acid binding. Biochemistry. 1971 May 25;10(11):2094–2100. doi: 10.1021/bi00787a021. [DOI] [PubMed] [Google Scholar]
- Morgan R. W., Christman M. F., Jacobson F. S., Storz G., Ames B. N. Hydrogen peroxide-inducible proteins in Salmonella typhimurium overlap with heat shock and other stress proteins. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8059–8063. doi: 10.1073/pnas.83.21.8059. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Neidhardt F. C., VanBogelen R. A., Vaughn V. The genetics and regulation of heat-shock proteins. Annu Rev Genet. 1984;18:295–329. doi: 10.1146/annurev.ge.18.120184.001455. [DOI] [PubMed] [Google Scholar]
- O'Farrell P. Z., Goodman H. M., O'Farrell P. H. High resolution two-dimensional electrophoresis of basic as well as acidic proteins. Cell. 1977 Dec;12(4):1133–1141. doi: 10.1016/0092-8674(77)90176-3. [DOI] [PubMed] [Google Scholar]
- Ozaki M., Wada A., Fujita N., Ishihama A. Growth phase-dependent modification of RNA polymerase in Escherichia coli. Mol Gen Genet. 1991 Nov;230(1-2):17–23. doi: 10.1007/BF00290644. [DOI] [PubMed] [Google Scholar]
- Perry R. D., Brubaker R. R. Transport of Ca2+ by Yersinia pestis. J Bacteriol. 1987 Oct;169(10):4861–4864. doi: 10.1128/jb.169.10.4861-4864.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Perry R. D., Harmon P. A., Bowmer W. S., Straley S. C. A low-Ca2+ response operon encodes the V antigen of Yersinia pestis. Infect Immun. 1986 Nov;54(2):428–434. doi: 10.1128/iai.54.2.428-434.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pollack C., Straley S. C., Klempner M. S. Probing the phagolysosomal environment of human macrophages with a Ca2+-responsive operon fusion in Yersinia pestis. 1986 Aug 28-Sep 3Nature. 322(6082):834–836. doi: 10.1038/322834a0. [DOI] [PubMed] [Google Scholar]
- Portnoy D. A., Moseley S. L., Falkow S. Characterization of plasmids and plasmid-associated determinants of Yersinia enterocolitica pathogenesis. Infect Immun. 1981 Feb;31(2):775–782. doi: 10.1128/iai.31.2.775-782.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Price S. B., Cowan C., Perry R. D., Straley S. C. The Yersinia pestis V antigen is a regulatory protein necessary for Ca2(+)-dependent growth and maximal expression of low-Ca2+ response virulence genes. J Bacteriol. 1991 Apr;173(8):2649–2657. doi: 10.1128/jb.173.8.2649-2657.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Price S. B., Leung K. Y., Barve S. S., Straley S. C. Molecular analysis of lcrGVH, the V antigen operon of Yersinia pestis. J Bacteriol. 1989 Oct;171(10):5646–5653. doi: 10.1128/jb.171.10.5646-5653.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Price S. B., Straley S. C. lcrH, a gene necessary for virulence of Yersinia pestis and for the normal response of Y. pestis to ATP and calcium. Infect Immun. 1989 May;57(5):1491–1498. doi: 10.1128/iai.57.5.1491-1498.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Protsenko O. A., Anisimov P. I., Mozharov O. T., Konnov N. P., Popov Iu A. Vyiavlenie i kharakteristika plazmid chumnogo mikroba, determiniruiushchikh sintez pestitsina I, antigena fraktsiia I i ékzotoksina "myshinogo" toksina. Genetika. 1983 Jul;19(7):1081–1090. [PubMed] [Google Scholar]
- Rosqvist R., Forsberg A., Rimpiläinen M., Bergman T., Wolf-Watz H. The cytotoxic protein YopE of Yersinia obstructs the primary host defence. Mol Microbiol. 1990 Apr;4(4):657–667. doi: 10.1111/j.1365-2958.1990.tb00635.x. [DOI] [PubMed] [Google Scholar]
- Sample A. K., Brubaker R. R. Post-translational regulation of Lcr plasmid-mediated peptides in pesticinogenic Yersinia pestis. Microb Pathog. 1987 Oct;3(4):239–248. doi: 10.1016/0882-4010(87)90057-x. [DOI] [PubMed] [Google Scholar]
- Sample A. K., Fowler J. M., Brubaker R. R. Modulation of the low-calcium response in Yersinia pestis via plasmid-plasmid interaction. Microb Pathog. 1987 Jun;2(6):443–453. doi: 10.1016/0882-4010(87)90051-9. [DOI] [PubMed] [Google Scholar]
- Sato K., Nakajima R., Hara F., Une T., Osada Y. Preparation of monoclonal antibody to V antigen from Yersinia pestis. Contrib Microbiol Immunol. 1991;12:225–229. [PubMed] [Google Scholar]
- Siegele D. A., Kolter R. Life after log. J Bacteriol. 1992 Jan;174(2):345–348. doi: 10.1128/jb.174.2.345-348.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Silver S., Toth K., Scribner H. Facilitated transport of calcium by cells and subcellular membranes of Bacillus subtilis and Escherichia coli. J Bacteriol. 1975 Jun;122(3):880–885. doi: 10.1128/jb.122.3.880-885.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sodeinde O. A., Goguen J. D. Genetic analysis of the 9.5-kilobase virulence plasmid of Yersinia pestis. Infect Immun. 1988 Oct;56(10):2743–2748. doi: 10.1128/iai.56.10.2743-2748.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sodeinde O. A., Sample A. K., Brubaker R. R., Goguen J. D. Plasminogen activator/coagulase gene of Yersinia pestis is responsible for degradation of plasmid-encoded outer membrane proteins. Infect Immun. 1988 Oct;56(10):2749–2752. doi: 10.1128/iai.56.10.2749-2752.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Storz G., Tartaglia L. A., Ames B. N. Transcriptional regulator of oxidative stress-inducible genes: direct activation by oxidation. Science. 1990 Apr 13;248(4952):189–194. doi: 10.1126/science.2183352. [DOI] [PubMed] [Google Scholar]
- Straley S. C., Brubaker R. R. Cytoplasmic and membrane proteins of yersiniae cultivated under conditions simulating mammalian intracellular environment. Proc Natl Acad Sci U S A. 1981 Feb;78(2):1224–1228. doi: 10.1073/pnas.78.2.1224. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Une T., Brubaker R. R. Roles of V antigen in promoting virulence and immunity in yersiniae. J Immunol. 1984 Oct;133(4):2226–2230. [PubMed] [Google Scholar]
- Une T., Nakajima R., Brubaker R. R. Roles of V antigen in promoting virulence in Yersiniae. Contrib Microbiol Immunol. 1987;9:179–185. [PubMed] [Google Scholar]
- Van Dyk T. K., Gatenby A. A., LaRossa R. A. Demonstration by genetic suppression of interaction of GroE products with many proteins. Nature. 1989 Nov 23;342(6248):451–453. doi: 10.1038/342451a0. [DOI] [PubMed] [Google Scholar]
- Winterbourn C. C., Hawkins R. E., Brian M., Carrell R. W. The estimation of red cell superoxide dismutase activity. J Lab Clin Med. 1975 Feb;85(2):337–341. [PubMed] [Google Scholar]
- Zahorchak R. J., Brubaker R. R. Effect of exogenous nucleotides on Ca2+ dependence and V antigen synthesis in Yersinia pestis. Infect Immun. 1982 Dec;38(3):953–959. doi: 10.1128/iai.38.3.953-959.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zahorchak R. J., Charnetzky W. T., Little R. V., Brubaker R. R. Consequences of Ca2+ deficiency on macromolecular synthesis and adenylate energy charge in Yersinia pestis. J Bacteriol. 1979 Sep;139(3):792–799. doi: 10.1128/jb.139.3.792-799.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]