Logo of iaiPermissionsJournals.ASM.orgJournalIAI ArticleJournal InfoAuthorsReviewers
Infect Immun. Dec 1994; 62(12): 5234–5241.
PMCID: PMC303259

Physiological basis of the low calcium response in Yersinia pestis.

Abstract

It is established that duplication in vitro of that amount of Ca2+ (2.5 mM) and Mg2+ (1.5 mM) present in blood permits vegetative growth of Yersinia pestis with repression of virulence factors encoded by the Lcr plasmid (Lcr+); similar simulation of intracellular fluid (no Ca2+ and 20 mM Mg2+) promotes bacteriostasis with induction of these virulence determinants. However, proliferation of yersiniae in mice occurs primarily within necrotic focal lesions (supplied by Ca(2+)-deficient host cell cytoplasm) within visceral organs rather than in Ca(2+)-sufficient blood. The present study addressed this enigma by defining conditions necessary for achieving vegetative growth of Lcr+ yersiniae at 37 degrees C in simulated intracellular fluid. Maximum optical densities were increased by substitution of K+ for Na+ and elimination of Cl-; the combination of Na+ plus L-glutamate was selectively toxic to Lcr+ cells. This phenomenon was attributed in part to the absence of aspartase in Y. pestis (a lesion known to facilitate massive accumulation of L-aspartate via transamination of the oxalacetate pool by L-glutamate). Replacement of L-glutamate by exogenous L-aspartate or alpha-ketoglutarate reversed this toxicity by favoring retention of oxalacetate. Proliferation of Lcr+ cells in a medium containing K+ and L-aspartate but lacking added Ca2+ and Na+ was markedly enhanced by increasing the concentration of fermentable carbohydrate. Accordingly, in the worst-case scenario (i.e., added Na+, Cl-, and L-glutamate), Lcr+ yersiniae underwent restriction of growth after one doubling, and in the best-case scenario (i.e., added K+ and L-aspartate), the organisms completed more than five doublings, thereby achieving full-scale growth. Both of these Ca(2+)-deficient media promoted maximum induction of Mg(2+)-induced V antigen, a virulence factor encoded by the Lcr plasmid.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.7M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Autenrieth IB, Vogel U, Preger S, Heymer B, Heesemann J. Experimental Yersinia enterocolitica infection in euthymic and T-cell-deficient athymic nude C57BL/6 mice: comparison of time course, histomorphology, and immune response. Infect Immun. 1993 Jun;61(6):2585–2595. [PMC free article] [PubMed]
  • Barve SS, Straley SC. lcrR, a low-Ca2(+)-response locus with dual Ca2(+)-dependent functions in Yersinia pestis. J Bacteriol. 1990 Aug;172(8):4661–4671. [PMC free article] [PubMed]
  • BAUGH CL, LANHAM JW, SURGALLA MJ. EFFECTS OF BICARBONATE ON GROWTH OF PASTEURELLA PESTIS. II. CARBON DIOXIDE FIXATION INTO OXALACETATE BY CELL-FREE EXTRACTS. J Bacteriol. 1964 Sep;88:553–558. [PMC free article] [PubMed]
  • Beesley ED, Brubaker RR, Janssen WA, Surgalla MJ. Pesticins. 3. Expression of coagulase and mechanism of fibrinolysis. J Bacteriol. 1967 Jul;94(1):19–26. [PMC free article] [PubMed]
  • Ben-Gurion R, Shafferman A. Essential virulence determinants of different Yersinia species are carried on a common plasmid. Plasmid. 1981 Mar;5(2):183–187. [PubMed]
  • 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. [PMC free article] [PubMed]
  • Bölin I, Wolf-Watz H. The plasmid-encoded Yop2b protein of Yersinia pseudotuberculosis is a virulence determinant regulated by calcium and temperature at the level of transcription. Mol Microbiol. 1988 Mar;2(2):237–245. [PubMed]
  • Brubaker RR. Growth of Pasteurella pseudotuberculosis in simulated intracellular and extracellular environments. J Infect Dis. 1967 Dec;117(5):403–417. [PubMed]
  • Brubaker RR. Interconversion of Purine Mononucleotides in Pasteurella pestis. Infect Immun. 1970 May;1(5):446–454. [PMC free article] [PubMed]
  • Brubaker RR. The genus Yersinia: biochemistry and genetics of virulence. Curr Top Microbiol Immunol. 1972;57:111–158. [PubMed]
  • Brubaker RR. Factors promoting acute and chronic diseases caused by yersiniae. Clin Microbiol Rev. 1991 Jul;4(3):309–324. [PMC free article] [PubMed]
  • Brubaker RR, Sample AK, Yu DZ, Zahorchak RJ, Hu PC, Fowler JM. Proteolysis of V antigen from Yersinia pestis. Microb Pathog. 1987 Jan;2(1):49–62. [PubMed]
  • BRUBAKER RR, SURGALLA MJ. 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. [PubMed]
  • Carter PB, Zahorchak RJ, Brubaker RR. Plague virulence antigens from Yersinia enterocolitica. Infect Immun. 1980 May;28(2):638–640. [PMC free article] [PubMed]
  • Charnetzky WT, Brubaker RR. RNA synthesis in Yersinia pestis during growth restriction in calcium-deficient medium. J Bacteriol. 1982 Mar;149(3):1089–1095. [PMC free article] [PubMed]
  • Cornelis GR, Biot T, Lambert de Rouvroit C, Michiels T, Mulder B, Sluiters C, Sory MP, Van Bouchaute M, Vanooteghem JC. The Yersinia yop regulon. Mol Microbiol. 1989 Oct;3(10):1455–1459. [PubMed]
  • CRUMPTON MJ, DAVIES DA. 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. [PubMed]
  • DELWICHE EA, FUKUI GM, ANDREWS AW, SURGALLA MJ. Environmental conditions affecting the population dynamics and the retention of virulence of Pasteurella pestis: the role of carbon dioxide. J Bacteriol. 1959 Mar;77(3):355–360. [PMC free article] [PubMed]
  • Dreyfus LA, Brubaker RR. Consequences of aspartase deficiency in Yersinia pestis. J Bacteriol. 1978 Nov;136(2):757–764. [PMC free article] [PubMed]
  • Ferber DM, Brubaker RR. Plasmids in Yersinia pestis. Infect Immun. 1981 Feb;31(2):839–841. [PMC free article] [PubMed]
  • Fetherston JD, Schuetze P, Perry RD. Loss of the pigmentation phenotype in Yersinia pestis is due to the spontaneous deletion of 102 kb of chromosomal DNA which is flanked by a repetitive element. Mol Microbiol. 1992 Sep;6(18):2693–2704. [PubMed]
  • FUKUI GM, OGG JE, WESSMAN GE, SURGALLA MJ. Studies on the relation of cultural conditions and virulence of Pasteurella pestis. J Bacteriol. 1957 Dec;74(6):714–717. [PMC free article] [PubMed]
  • Galyov EE, Håkansson S, Forsberg A, Wolf-Watz H. A secreted protein kinase of Yersinia pseudotuberculosis is an indispensable virulence determinant. Nature. 1993 Feb 25;361(6414):730–732. [PubMed]
  • Goguen JD, Yother J, Straley SC. Genetic analysis of the low calcium response in Yersinia pestis mu d1(Ap lac) insertion mutants. J Bacteriol. 1984 Dec;160(3):842–848. [PMC free article] [PubMed]
  • Gowrishankar J. Identification of osmoresponsive genes in Escherichia coli: evidence for participation of potassium and proline transport systems in osmoregulation. J Bacteriol. 1985 Oct;164(1):434–445. [PMC free article] [PubMed]
  • Håkansson S, Bergman T, Vanooteghem JC, Cornelis G, Wolf-Watz H. YopB and YopD constitute a novel class of Yersinia Yop proteins. Infect Immun. 1993 Jan;61(1):71–80. [PMC free article] [PubMed]
  • HIGUCHI K, KUPFERBERG LL, SMITH JL. Studies on the nutrition and physiology of Pasteurella pestis. III. Effects of calcium ions on the growth of virulent and avirulent strains of Pasteurella pestis. J Bacteriol. 1959 Mar;77(3):317–321. [PMC free article] [PubMed]
  • HIGUCHI K, SMITH JL. Studies on the nutrition and physiology of Pasteurella pestis. VI. A differential plating medium for the estimation of the mutation rate to avirulence. J Bacteriol. 1961 Apr;81:605–608. [PMC free article] [PubMed]
  • Hillier S, Charnetzky WT. Glyoxylate bypass enzymes in Yersinia species and multiple forms of isocitrate lyase in Yersinia pestis. J Bacteriol. 1981 Jan;145(1):452–458. [PMC free article] [PubMed]
  • KUPFERBERG LL, HIGUCHI K. Role of calcium ions in the stimulation of growth of virulent strains of Pasteurella pestis. J Bacteriol. 1958 Jul;76(1):120–121. [PMC free article] [PubMed]
  • LAWTON WD, ERDMAN RL, SURGALLA MJ. BIOSYNTHESIS AND PURIFICATION OF V AND W ANTIGEN IN PASTEURELLA PESTIS. J Immunol. 1963 Aug;91:179–184. [PubMed]
  • LOWRY OH, ROSEBROUGH NJ, FARR AL, RANDALL RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed]
  • Lucier TS, Brubaker RR. 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. [PMC free article] [PubMed]
  • Mehigh RJ, Braubaker RR. Major stable peptides of Yersinia pestis synthesized during the low-calcium response. Infect Immun. 1993 Jan;61(1):13–22. [PMC free article] [PubMed]
  • Mehigh RJ, Sample AK, Brubaker RR. Expression of the low calcium response in Yersinia pestis. Microb Pathog. 1989 Mar;6(3):203–217. [PubMed]
  • Nakajima R, Brubaker RR. Association between virulence of Yersinia pestis and suppression of gamma interferon and tumor necrosis factor alpha. Infect Immun. 1993 Jan;61(1):23–31. [PMC free article] [PubMed]
  • Portnoy DA, Falkow S. Virulence-associated plasmids from Yersinia enterocolitica and Yersinia pestis. J Bacteriol. 1981 Dec;148(3):877–883. [PMC free article] [PubMed]
  • Price SB, Cowan C, Perry RD, Straley SC. 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. [PMC free article] [PubMed]
  • Price SB, Straley SC. 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. [PMC free article] [PubMed]
  • 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. [PubMed]
  • Rosqvist R, Magnusson KE, Wolf-Watz H. Target cell contact triggers expression and polarized transfer of Yersinia YopE cytotoxin into mammalian cells. EMBO J. 1994 Feb 15;13(4):964–972. [PMC free article] [PubMed]
  • Simonet M, Richard S, Berche P. Electron microscopic evidence for in vivo extracellular localization of Yersinia pseudotuberculosis harboring the pYV plasmid. Infect Immun. 1990 Mar;58(3):841–845. [PMC free article] [PubMed]
  • Skryzpek E, Straley SC. LcrG, a secreted protein involved in negative regulation of the low-calcium response in Yersinia pestis. J Bacteriol. 1993 Jun;175(11):3520–3528. [PMC free article] [PubMed]
  • Straley SC, Brubaker RR. Localization in Yersinia pestis of peptides associated with virulence. Infect Immun. 1982 Apr;36(1):129–135. [PMC free article] [PubMed]
  • Straley SC, Cibull ML. Differential clearance and host-pathogen interactions of YopE- and YopK- YopL- Yersinia pestis in BALB/c mice. Infect Immun. 1989 Apr;57(4):1200–1210. [PMC free article] [PubMed]
  • Straley SC, Plano GV, Skrzypek E, Haddix PL, Fields KA. Regulation by Ca2+ in the Yersinia low-Ca2+ response. Mol Microbiol. 1993 Jun;8(6):1005–1010. [PubMed]
  • Straley SC, Skrzypek E, Plano GV, Bliska JB. Yops of Yersinia spp. pathogenic for humans. Infect Immun. 1993 Aug;61(8):3105–3110. [PMC free article] [PubMed]
  • Une T, Brubaker RR. In vivo comparison of avirulent Vwa- and Pgm- or Pstr phenotypes of yersiniae. Infect Immun. 1984 Mar;43(3):895–900. [PMC free article] [PubMed]
  • Une T, Brubaker RR. Roles of V antigen in promoting virulence and immunity in yersiniae. J Immunol. 1984 Oct;133(4):2226–2230. [PubMed]
  • Une T, Nakajima R, Brubaker RR. Roles of V antigen in promoting virulence in Yersiniae. Contrib Microbiol Immunol. 1987;9:179–185. [PubMed]
  • WESSMAN GE, MILLER DJ, SURGALLA MJ. Toxic effect of glucose on virulent Pasteurella pestis in chemically defined media. J Bacteriol. 1958 Oct;76(4):368–375. [PMC free article] [PubMed]
  • Yother J, Goguen JD. Isolation and characterization of Ca2+-blind mutants of Yersinia pestis. J Bacteriol. 1985 Nov;164(2):704–711. [PMC free article] [PubMed]
  • Zahorchak RJ, Brubaker RR. Effect of exogenous nucleotides on Ca2+ dependence and V antigen synthesis in Yersinia pestis. Infect Immun. 1982 Dec;38(3):953–959. [PMC free article] [PubMed]
  • Zahorchak RJ, Charnetzky WT, Little RV, Brubaker RR. Consequences of Ca2+ deficiency on macromolecular synthesis and adenylate energy charge in Yersinia pestis. J Bacteriol. 1979 Sep;139(3):792–799. [PMC free article] [PubMed]

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

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...