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PMID: 1657887 Published · ppublish English Comparative Study Journal Article Research Support, U.S. Gov't, P.H.S.

LcrD, a membrane-bound regulator of the Yersinia pestis low-calcium response.

Journal of bacteriology ·Vol. 173 ·No. 22 ·1991-11-00 ·Pages 7293-303

Plano GV, Barve SS, Straley SC

Abstract

Yersinia pestis, the etiologic agent of bubonic plague, contains a 75-kb virulence plasmid, called pCD1 in Y. pestis KIM. The low-Ca(2+)-response genes of Y. pestis regulate both bacterial growth and the expression of pCD1-encoded virulence determinants in response to temperature and the presence of Ca2+ or nucleotides. This study characterizes the nucleotide sequence and protein product of the lcrD locus. An lcrD mutant, in contrast to the parent Y. pestis, did not undergo growth restriction or induce strong expression of the V antigen when grown under conditions (37 degrees C, no Ca2+) expected to elicit maximal expression of pCD1 genes. DNA sequence analysis of the cloned lcrD locus showed a single open reading frame that could encode a protein with a molecular weight of 77,804 and a pI of 4.88. LcrD was identified as a 70-kDa inner membrane protein by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblot analysis. LcrD membrane topology was investigated by using lcrD-phoA translational fusions generated with the transposon TnphoA. The alkaline phosphatase activities of the resultant hybrid proteins were consistent with a model predicting eight amino-terminal transmembrane segments that anchor a large cytoplasmic carboxyl-terminal domain to the inner membrane.

MeSH Terms
Alkaline Phosphatase/genetics,metabolism Amino Acid Sequence Bacterial Proteins/genetics,isolation & purification Base Sequence Calcium/pharmacology Cell Membrane/ultrastructure Cloning, Molecular DNA Transposable Elements Escherichia coli/genetics Membrane Proteins/genetics,isolation & purification Models, Structural Molecular Sequence Data Mutagenesis, Insertional Open Reading Frames Plasmids Protein Conformation Recombinant Fusion Proteins/metabolism Restriction Mapping Sequence Homology, Nucleic Acid Transcription, Genetic Virulence/genetics Yersinia pestis/drug effects,genetics,pathogenicity,physiology
Chemicals
Bacterial Proteins DNA Transposable Elements Membrane Proteins Recombinant Fusion Proteins LcrD protein, Yersinia Alkaline Phosphatase Calcium
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Plano G V
Department of Microbiology and Immunology, Chandler Medical Center, University of Kentucky, Lexington 40536.
Barve S S
Straley S C
References (74)
74 references, click to expand
  1. Determinants of membrane protein topology.
    Proc Natl Acad Sci U S A. 1987 Dec;84(23):8525-9 PMID: 3317413
  2. Identification and mapping of the temperature-inducible, plasmid-encoded proteins of Yersinia spp.
    Infect Immun. 1988 Feb;56(2):343-8 PMID: 3338844
  3. Optimal alignments in linear space.
    Comput Appl Biosci. 1988 Mar;4(1):11-7 PMID: 3382986
  4. A genetic approach to analyzing membrane protein topology.
    Science. 1986 Sep 26;233(4771):1403-8 PMID: 3529391
  5. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes.
    J Mol Biol. 1986 May 5;189(1):113-30 PMID: 3537305
  6. Transport of Ca2+ by Yersinia pestis.
    J Bacteriol. 1987 Oct;169(10):4861-4 PMID: 3654586
  7. Regulation of expression of V antigen and outer membrane proteins in Yersinia pestis.
    Contrib Microbiol Immunol. 1987;9:173-8 PMID: 3665495
  8. Roles of V antigen in promoting virulence in Yersiniae.
    Contrib Microbiol Immunol. 1987;9:179-85 PMID: 3665496
  9. The detection and classification of membrane-spanning proteins.
    Biochim Biophys Acta. 1985 May 28;815(3):468-76 PMID: 3838905
  10. Temperature-controlled plasmid regulon associated with low calcium response in Yersinia pestis.
    J Bacteriol. 1986 Feb;165(2):443-7 PMID: 3944056
  11. Characterization of plasmids and plasmid-associated determinants of Yersinia enterocolitica pathogenesis.
    Infect Immun. 1981 Feb;31(2):775-82 PMID: 7216474
  12. Plasmids in Yersinia pestis.
    Infect Immun. 1981 Feb;31(2):839-41 PMID: 7216478
  13. Essential virulence determinants of different Yersinia species are carried on a common plasmid.
    Plasmid. 1981 Mar;5(2):183-7 PMID: 7243971
  14. The basis of virulence in Pasteurella pestis: an antigen determining virulence.
    Br J Exp Pathol. 1956 Oct;37(5):481-93 PMID: 13374206
  15. lcrR, a low-Ca2(+)-response locus with dual Ca2(+)-dependent functions in Yersinia pestis.
    J Bacteriol. 1990 Aug;172(8):4661-71 PMID: 1695896
  16. Tyrosine phosphate hydrolysis of host proteins by an essential Yersinia virulence determinant.
    Proc Natl Acad Sci U S A. 1991 Feb 15;88(4):1187-91 PMID: 1705028
  17. 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):1607-16 PMID: 1705541
  18. Timing of flagellar gene expression in the Caulobacter cell cycle is determined by a transcriptional cascade of positive regulatory genes.
    J Bacteriol. 1991 Feb;173(4):1514-22 PMID: 1847367
  19. BIOSYNTHESIS AND PURIFICATION OF V AND W ANTIGEN IN PASTEURELLA PESTIS.
    J Immunol. 1963 Aug;91:179-84 PMID: 14061374
  20. The distribution of positively charged residues in bacterial inner membrane proteins correlates with the trans-membrane topology.
    EMBO J. 1986 Nov;5(11):3021-7 PMID: 16453726
  21. High resolution two-dimensional electrophoresis of basic as well as acidic proteins.
    Cell. 1977 Dec;12(4):1133-41 PMID: 23215
  22. Plasmids of Escherichia coli as cloning vectors.
    Methods Enzymol. 1979;68:245-67 PMID: 232214
  23. High resolution two-dimensional electrophoresis of proteins.
    J Biol Chem. 1975 May 25;250(10):4007-21 PMID: 236308
  24. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  25. A rapid alkaline extraction procedure for screening recombinant plasmid DNA.
    Nucleic Acids Res. 1979 Nov 24;7(6):1513-23 PMID: 388356
  26. Consequences of Ca2+ deficiency on macromolecular synthesis and adenylate energy charge in Yersinia pestis.
    J Bacteriol. 1979 Sep;139(3):792-9 PMID: 479109
  27. Bacteriophage T7 early promoters: nucleotide sequences of two RNA polymerase binding sites.
    J Mol Biol. 1975 Dec 15;99(3):419-43 PMID: 765476
  28. Nucleotide sequence of an RNA polymerase binding site at an early T7 promoter.
    Proc Natl Acad Sci U S A. 1975 Mar;72(3):784-8 PMID: 1093168
  29. Analysis of virC, an operon involved in the secretion of Yop proteins by Yersinia enterocolitica.
    J Bacteriol. 1991 Aug;173(16):4994-5009 PMID: 1860816
  30. 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):2649-57 PMID: 1901573
  31. Identification and initial topological analysis of the Rickettsia prowazekii ATP/ADP translocase.
    J Bacteriol. 1991 Jun;173(11):3389-96 PMID: 1904433
  32. 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):7283-92 PMID: 1938923
  33. The lcrE gene is part of an operon in the lcr region of Yersinia enterocolitica O:3.
    J Bacteriol. 1990 Jun;172(6):3152-62 PMID: 2160939
  34. lac permease of Escherichia coli: topology and sequence elements promoting membrane insertion.
    Proc Natl Acad Sci U S A. 1990 Jul;87(13):4937-41 PMID: 2164211
  35. Protein tyrosine phosphatase activity of an essential virulence determinant in Yersinia.
    Science. 1990 Aug 3;249(4968):553-6 PMID: 2166336
  36. The cytotoxic protein YopE of Yersinia obstructs the primary host defence.
    Mol Microbiol. 1990 Apr;4(4):657-67 PMID: 2191183
  37. Identification and cloning of a hemin storage locus involved in the pigmentation phenotype of Yersinia pestis.
    J Bacteriol. 1990 Oct;172(10):5929-37 PMID: 2211518
  38. A highly efficient electroporation system for transformation of Yersinia.
    Gene. 1990 Mar 1;87(1):133-7 PMID: 2332165
  39. YopM inhibits platelet aggregation and is necessary for virulence of Yersinia pestis in mice.
    Infect Immun. 1990 Oct;58(10):3262-71 PMID: 2401564
  40. Molecular analysis of lcrGVH, the V antigen operon of Yersinia pestis.
    J Bacteriol. 1989 Oct;171(10):5646-53 PMID: 2477361
  41. Expression of the temperature-inducible outer membrane proteins of yersiniae.
    Infect Immun. 1985 Apr;48(1):234-40 PMID: 3980086
  42. Mechanism of assembly of the outer membrane of Salmonella typhimurium. Isolation and characterization of cytoplasmic and outer membrane.
    J Biol Chem. 1972 Jun 25;247(12):3962-72 PMID: 4555955
  43. 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):1342-6 PMID: 4598299
  44. Genetic analysis of the low calcium response in Yersinia pestis mu d1(Ap lac) insertion mutants.
    J Bacteriol. 1984 Dec;160(3):842-8 PMID: 6094509
  45. Identification of the secY (prlA) gene product involved in protein export in Escherichia coli.
    Mol Gen Genet. 1984;197(2):204-8 PMID: 6097791
  46. Prediction of protein antigenic determinants from amino acid sequences.
    Proc Natl Acad Sci U S A. 1981 Jun;78(6):3824-8 PMID: 6167991
  47. The Vwa+ virulence factor of yersiniae: the molecular basis of the attendant nutritional requirement for Ca++.
    Rev Infect Dis. 1983 Sep-Oct;5 Suppl 4:S748-58 PMID: 6195719
  48. New M13 vectors for cloning.
    Methods Enzymol. 1983;101:20-78 PMID: 6310323
  49. Mud(Ap, lac)-generated fusions in studies of gene expression.
    Methods Enzymol. 1983;100:501-9 PMID: 6312264
  50. Characterization of common virulence plasmids in Yersinia species and their role in the expression of outer membrane proteins.
    Infect Immun. 1984 Jan;43(1):108-14 PMID: 6317562
  51. Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis.
    Gene. 1983 Dec;26(1):101-6 PMID: 6323249
  52. Compilation and analysis of Escherichia coli promoter DNA sequences.
    Nucleic Acids Res. 1983 Apr 25;11(8):2237-55 PMID: 6344016
  53. In vivo comparison of avirulent Vwa- and Pgm- or Pstr phenotypes of yersiniae.
    Infect Immun. 1984 Mar;43(3):895-900 PMID: 6365786
  54. Roles of V antigen in promoting virulence and immunity in yersiniae.
    J Immunol. 1984 Oct;133(4):2226-30 PMID: 6381597
  55. Sequence of the lactose permease gene.
    Nature. 1980 Feb 7;283(5747):541-5 PMID: 6444453
  56. Cytoplasmic and membrane proteins of yersiniae cultivated under conditions simulating mammalian intracellular environment.
    Proc Natl Acad Sci U S A. 1981 Feb;78(2):1224-8 PMID: 6940138
  57. A simple method for displaying the hydropathic character of a protein.
    J Mol Biol. 1982 May 5;157(1):105-32 PMID: 7108955
  58. Effect of exogenous nucleotides on Ca2+ dependence and V antigen synthesis in Yersinia pestis.
    Infect Immun. 1982 Dec;38(3):953-9 PMID: 7152680
  59. Identification of additional virulence determinants on the pYV plasmid of Yersinia enterocolitica W227.
    Infect Immun. 1989 Aug;57(8):2534-41 PMID: 2545628
  60. Protein phosphorylation and regulation of adaptive responses in bacteria.
    Microbiol Rev. 1989 Dec;53(4):450-90 PMID: 2556636
  61. 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):4623-32 PMID: 2670888
  62. 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):1491-8 PMID: 2707857
  63. The use of transposon TnphoA to detect genes for cell envelope proteins subject to a common regulatory stimulus. Analysis of osmotically regulated genes in Escherichia coli.
    J Mol Biol. 1987 May 20;195(2):289-97 PMID: 2821274
  64. Plasminogen activator/coagulase gene of Yersinia pestis is responsible for degradation of plasmid-encoded outer membrane proteins.
    Infect Immun. 1988 Oct;56(10):2749-52 PMID: 2843471
  65. Genetic analysis of the plasmid region controlling virulence in Yersinia enterocolitica 0:9 by Mini-Mu insertions and lac gene fusions.
    Microb Pathog. 1986 Aug;1(4):349-59 PMID: 2854600
  66. Differential clearance and host-pathogen interactions of YopE- and YopK- YopL- Yersinia pestis in BALB/c mice.
    Infect Immun. 1989 Apr;57(4):1200-10 PMID: 2925246
  67. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors.
    Gene. 1985;33(1):103-19 PMID: 2985470
  68. Isolation and characterization of Ca2+-blind mutants of Yersinia pestis.
    J Bacteriol. 1985 Nov;164(2):704-11 PMID: 2997127
  69. TnphoA: a transposon probe for protein export signals.
    Proc Natl Acad Sci U S A. 1985 Dec;82(23):8129-33 PMID: 2999794
  70. 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):445-54 PMID: 3002984
  71. A low-Ca2+ response operon encodes the V antigen of Yersinia pestis.
    Infect Immun. 1986 Nov;54(2):428-34 PMID: 3021629
  72. The plasmid-encoded outer-membrane proteins of Yersinia pestis.
    Rev Infect Dis. 1988 Jul-Aug;10 Suppl 2:S323-6 PMID: 3055200
  73. A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes.
    Proc Natl Acad Sci U S A. 1985 Feb;82(4):1074-8 PMID: 3156376
  74. Improved tools for biological sequence comparison.
    Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444-8 PMID: 3162770
Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.173.22.7293-7303.1991
Published
1991-11-00
Pages
7293-303
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC209237
Subset
IM
Grants
NIAID NIH HHS · AI21017 · United States
NCI NIH HHS · CA09509-06 · United States
Databases
GENBANK
M63255, M64326, M64327, M64328, M64329, M64330, M64331, M64332, M77014, S65012
External Links
PubMed source DOI: 10.1128/jb.173.22.7293-7303.1991 PMC Full Text View journal details
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