• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jbacterPermissionsJournals.ASM.orgJournalJB ArticleJournal InfoAuthorsReviewers
J Bacteriol. Feb 1994; 176(3): 569–579.
PMCID: PMC205092

A low-Ca2+ response (LCR) secretion (ysc) locus lies within the lcrB region of the LCR plasmid in Yersinia pestis.

Abstract

The causative agent of plague, Yersinia pestis, contains a 75-kb plasmid, pCD1, which carries a virulence-related stimulon called the low-Ca2+ response stimulon (LCRS). LCRS operons are regulated by the environmental signals of temperature and Ca2+. This study characterized a portion of the lcrB region of pCD1, known to contain at least one gene necessary for the regulation of LCRS operons by Ca2+. The sequence of a 2-kb region revealed three open reading frames, designated yscQ, yscR, and yscS, predicted to encode acidic proteins of 34.4, 24.4, and 8.5 kDa. All three proteins were homologous to proteins involved in flagellar function or virulence. An antipeptide antibody specific for YscR was used to localize YscR to the inner membrane of Y. pestis. Analysis of yscR-phoA fusions supported a model for yscR which predicts four transmembrane regions and a large, central hydrophilic domain. In-frame deletion mutations of yscQ and yscR were constructed and moved into Y. pestis. Both mutants failed to show the restriction of growth that normally accompanies maximal LCRS induction. Unlike the parent Y. pestis, the yscR mutant did not respond to the absence of Ca2+ by increasing the net transcription or translation of the LCRS-encoded V antigen, YopM, or LcrG. The yscR mutant also was defective for secretion of V antigen, YopM, and LcrG. These findings implicate a dual role for YscR in regulation of LCRS operons and secretion of LCRS proteins and add to the developing picture of how secretion of virulence proteins may be coupled to transcriptional regulation in yersiniae.

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 (2.3M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Allaoui A, Sansonetti PJ, Parsot C. MxiJ, a lipoprotein involved in secretion of Shigella Ipa invasins, is homologous to YscJ, a secretion factor of the Yersinia Yop proteins. J Bacteriol. 1992 Dec;174(23):7661–7669. [PMC free article] [PubMed]
  • Andrews GP, Maurelli AT. mxiA of Shigella flexneri 2a, which facilitates export of invasion plasmid antigens, encodes a homolog of the low-calcium-response protein, LcrD, of Yersinia pestis. Infect Immun. 1992 Aug;60(8):3287–3295. [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]
  • 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]
  • Bischoff DS, Ordal GW. Identification and characterization of FliY, a novel component of the Bacillus subtilis flagellar switch complex. Mol Microbiol. 1992 Sep;6(18):2715–2723. [PubMed]
  • Bischoff DS, Weinreich MD, Ordal GW. Nucleotide sequences of Bacillus subtilis flagellar biosynthetic genes fliP and fliQ and identification of a novel flagellar gene, fliZ. J Bacteriol. 1992 Jun;174(12):4017–4025. [PMC free article] [PubMed]
  • Bölin I, Portnoy DA, Wolf-Watz H. Expression of the temperature-inducible outer membrane proteins of yersiniae. Infect Immun. 1985 Apr;48(1):234–240. [PMC free article] [PubMed]
  • Brubaker RR. The V antigen of yersiniae: an overview. Contrib Microbiol Immunol. 1991;12:127–133. [PubMed]
  • BURROWS TW. An antigen determining virulence in Pasteurella pestis. Nature. 1956 Mar 3;177(4505):426–427. [PubMed]
  • Cornelis G, Laroche Y, Balligand G, Sory MP, Wauters G. Yersinia enterocolitica, a primary model for bacterial invasiveness. Rev Infect Dis. 1987 Jan-Feb;9(1):64–87. [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]
  • Cornelis GR, Sluiters C, Delor I, Geib D, Kaniga K, Lambert de Rouvroit C, Sory MP, Vanooteghem JC, Michiels T. ymoA, a Yersinia enterocolitica chromosomal gene modulating the expression of virulence functions. Mol Microbiol. 1991 May;5(5):1023–1034. [PubMed]
  • Eisenberg D, Schwarz E, Komaromy M, Wall R. Analysis of membrane and surface protein sequences with the hydrophobic moment plot. J Mol Biol. 1984 Oct 15;179(1):125–142. [PubMed]
  • Fenselau S, Balbo I, Bonas U. Determinants of pathogenicity in Xanthomonas campestris pv. vesicatoria are related to proteins involved in secretion in bacterial pathogens of animals. Mol Plant Microbe Interact. 1992 Sep-Oct;5(5):390–396. [PubMed]
  • Forsberg A, Viitanen AM, Skurnik M, Wolf-Watz H. The surface-located YopN protein is involved in calcium signal transduction in Yersinia pseudotuberculosis. Mol Microbiol. 1991 Apr;5(4):977–986. [PubMed]
  • Galán JE, Ginocchio C, Costeas P. Molecular and functional characterization of the Salmonella invasion gene invA: homology of InvA to members of a new protein family. J Bacteriol. 1992 Jul;174(13):4338–4349. [PMC free article] [PubMed]
  • Garnier J, Osguthorpe DJ, Robson B. Analysis of the accuracy and implications of simple methods for predicting the secondary structure of globular proteins. J Mol Biol. 1978 Mar 25;120(1):97–120. [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]
  • Gough CL, Genin S, Zischek C, Boucher CA. hrp genes of Pseudomonas solanacearum are homologous to pathogenicity determinants of animal pathogenic bacteria and are conserved among plant pathogenic bacteria. Mol Plant Microbe Interact. 1992 Sep-Oct;5(5):384–389. [PubMed]
  • Groisman EA, Ochman H. Cognate gene clusters govern invasion of host epithelial cells by Salmonella typhimurium and Shigella flexneri. EMBO J. 1993 Oct;12(10):3779–3787. [PMC free article] [PubMed]
  • Haddix PL, Straley SC. Structure and regulation of the Yersinia pestis yscBCDEF operon. J Bacteriol. 1992 Jul;174(14):4820–4828. [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]
  • Hsia RC, Small PL, Bavoil PM. Characterization of virulence genes of enteroinvasive Escherichia coli by TnphoA mutagenesis: identification of invX, a gene required for entry into HEp-2 cells. J Bacteriol. 1993 Aug;175(15):4817–4823. [PMC free article] [PubMed]
  • Hwang I, Lim SM, Shaw PD. Cloning and characterization of pathogenicity genes from Xanthomonas campestris pv. glycines. J Bacteriol. 1992 Mar;174(6):1923–1931. [PMC free article] [PubMed]
  • Irikura VM, Kihara M, Yamaguchi S, Sockett H, Macnab RM. Salmonella typhimurium fliG and fliN mutations causing defects in assembly, rotation, and switching of the flagellar motor. J Bacteriol. 1993 Feb;175(3):802–810. [PMC free article] [PubMed]
  • Jones CJ, Macnab RM. Flagellar assembly in Salmonella typhimurium: analysis with temperature-sensitive mutants. J Bacteriol. 1990 Mar;172(3):1327–1339. [PMC free article] [PubMed]
  • Kado CI, Liu ST. Rapid procedure for detection and isolation of large and small plasmids. J Bacteriol. 1981 Mar;145(3):1365–1373. [PMC free article] [PubMed]
  • Kihara M, Homma M, Kutsukake K, Macnab RM. Flagellar switch of Salmonella typhimurium: gene sequences and deduced protein sequences. J Bacteriol. 1989 Jun;171(6):3247–3257. [PMC free article] [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [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]
  • Leung KY, Reisner BS, Straley SC. YopM inhibits platelet aggregation and is necessary for virulence of Yersinia pestis in mice. Infect Immun. 1990 Oct;58(10):3262–3271. [PMC free article] [PubMed]
  • Leung KY, Straley SC. The yopM gene of Yersinia pestis encodes a released protein having homology with the human platelet surface protein GPIb alpha. J Bacteriol. 1989 Sep;171(9):4623–4632. [PMC free article] [PubMed]
  • Lindler LE, Klempner MS, Straley SC. 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. [PMC free article] [PubMed]
  • Malakooti J, Komeda Y, Matsumura P. DNA sequence analysis, gene product identification, and localization of flagellar motor components of Escherichia coli. J Bacteriol. 1989 May;171(5):2728–2734. [PMC free article] [PubMed]
  • Manoil C, Beckwith J. TnphoA: a transposon probe for protein export signals. Proc Natl Acad Sci U S A. 1985 Dec;82(23):8129–8133. [PMC free article] [PubMed]
  • Michiels T, Cornelis GR. Secretion of hybrid proteins by the Yersinia Yop export system. J Bacteriol. 1991 Mar;173(5):1677–1685. [PMC free article] [PubMed]
  • Michiels T, Vanooteghem JC, Lambert de Rouvroit C, China B, Gustin A, Boudry P, Cornelis GR. Analysis of virC, an operon involved in the secretion of Yop proteins by Yersinia enterocolitica. J Bacteriol. 1991 Aug;173(16):4994–5009. [PMC free article] [PubMed]
  • Michiels T, Wattiau P, Brasseur R, Ruysschaert JM, Cornelis G. Secretion of Yop proteins by Yersiniae. Infect Immun. 1990 Sep;58(9):2840–2849. [PMC free article] [PubMed]
  • Miller S, Pesci EC, Pickett CL. A Campylobacter jejuni homolog of the LcrD/FlbF family of proteins is necessary for flagellar biogenesis. Infect Immun. 1993 Jul;61(7):2930–2936. [PMC free article] [PubMed]
  • Miller VL, Mekalanos JJ. A novel suicide vector and its use in construction of insertion mutations: osmoregulation of outer membrane proteins and virulence determinants in Vibrio cholerae requires toxR. J Bacteriol. 1988 Jun;170(6):2575–2583. [PMC free article] [PubMed]
  • Mulholland V, Hinton JC, Sidebotham J, Toth IK, Hyman LJ, Pérombelon MC, Reeves PJ, Salmond GP. A pleiotropic reduced virulence (Rvi-) mutant of Erwinia carotovora subspecies atroseptica is defective in flagella assembly proteins that are conserved in plant and animal bacterial pathogens. Mol Microbiol. 1993 Jul;9(2):343–356. [PubMed]
  • Mullis KB, Faloona FA. Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. Methods Enzymol. 1987;155:335–350. [PubMed]
  • Myers EW, Miller W. Optimal alignments in linear space. Comput Appl Biosci. 1988 Mar;4(1):11–17. [PubMed]
  • Perry RD, Harmon PA, Bowmer WS, Straley SC. A low-Ca2+ response operon encodes the V antigen of Yersinia pestis. Infect Immun. 1986 Nov;54(2):428–434. [PMC free article] [PubMed]
  • Perry RD, Pendrak ML, Schuetze P. Identification and cloning of a hemin storage locus involved in the pigmentation phenotype of Yersinia pestis. J Bacteriol. 1990 Oct;172(10):5929–5937. [PMC free article] [PubMed]
  • Plano GV, Barve SS, Straley SC. LcrD, a membrane-bound regulator of the Yersinia pestis low-calcium response. J Bacteriol. 1991 Nov;173(22):7293–7303. [PMC free article] [PubMed]
  • Plano GV, Straley SC. Multiple effects of lcrD mutations in Yersinia pestis. J Bacteriol. 1993 Jun;175(11):3536–3545. [PMC free article] [PubMed]
  • Portnoy DA, Blank HF, Kingsbury DT, Falkow S. Genetic analysis of essential plasmid determinants of pathogenicity in Yersinia pestis. J Infect Dis. 1983 Aug;148(2):297–304. [PubMed]
  • Portnoy DA, Moseley SL, Falkow S. Characterization of plasmids and plasmid-associated determinants of Yersinia enterocolitica pathogenesis. Infect Immun. 1981 Feb;31(2):775–782. [PMC free article] [PubMed]
  • Price SB, Leung KY, Barve SS, Straley SC. Molecular analysis of lcrGVH, the V antigen operon of Yersinia pestis. J Bacteriol. 1989 Oct;171(10):5646–5653. [PMC free article] [PubMed]
  • Protsenko OA, Anisimov PI, Mozharov OT, Konnov NP, 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]
  • Pugsley AP. The complete general secretory pathway in gram-negative bacteria. Microbiol Rev. 1993 Mar;57(1):50–108. [PMC free article] [PubMed]
  • Ramakrishnan G, Zhao JL, Newton A. The cell cycle-regulated flagellar gene flbF of Caulobacter crescentus is homologous to a virulence locus (lcrD) of Yersinia pestis. J Bacteriol. 1991 Nov;173(22):7283–7292. [PMC free article] [PubMed]
  • Reisner BS, Straley SC. Yersinia pestis YopM: thrombin binding and overexpression. Infect Immun. 1992 Dec;60(12):5242–5252. [PMC free article] [PubMed]
  • Rosqvist R, Bölin I, Wolf-Watz H. Inhibition of phagocytosis in Yersinia pseudotuberculosis: a virulence plasmid-encoded ability involving the Yop2b protein. Infect Immun. 1988 Aug;56(8):2139–2143. [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, Forsberg A, Wolf-Watz H. Intracellular targeting of the Yersinia YopE cytotoxin in mammalian cells induces actin microfilament disruption. Infect Immun. 1991 Dec;59(12):4562–4569. [PMC free article] [PubMed]
  • Sanders LA, Van Way S, Mullin DA. Characterization of the Caulobacter crescentus flbF promoter and identification of the inferred FlbF product as a homolog of the LcrD protein from a Yersinia enterocolitica virulence plasmid. J Bacteriol. 1992 Feb;174(3):857–866. [PMC free article] [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PMC free article] [PubMed]
  • Sasakawa C, Komatsu K, Tobe T, Suzuki T, Yoshikawa M. Eight genes in region 5 that form an operon are essential for invasion of epithelial cells by Shigella flexneri 2a. J Bacteriol. 1993 Apr;175(8):2334–2346. [PMC free article] [PubMed]
  • Shine J, Dalgarno L. The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1342–1346. [PMC free article] [PubMed]
  • Skrzypek E, Haddix PL, Plano GV, Straley SC. New suicide vector for gene replacement in yersiniae and other gram-negative bacteria. Plasmid. 1993 Mar;29(2):160–163. [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]
  • Sodeinde OA, Goguen JD. Genetic analysis of the 9.5-kilobase virulence plasmid of Yersinia pestis. Infect Immun. 1988 Oct;56(10):2743–2748. [PMC free article] [PubMed]
  • Straley SC, Bowmer WS. Virulence genes regulated at the transcriptional level by Ca2+ in Yersinia pestis include structural genes for outer membrane proteins. Infect Immun. 1986 Feb;51(2):445–454. [PMC free article] [PubMed]
  • Straley SC, Brubaker RR. 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. [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]
  • 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]
  • Venkatesan MM, Buysse JM, Oaks EV. Surface presentation of Shigella flexneri invasion plasmid antigens requires the products of the spa locus. J Bacteriol. 1992 Mar;174(6):1990–2001. [PMC free article] [PubMed]
  • Yother J, Chamness TW, Goguen JD. Temperature-controlled plasmid regulon associated with low calcium response in Yersinia pestis. J Bacteriol. 1986 Feb;165(2):443–447. [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 Journal of Bacteriology 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...