Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1984 Jan;43(1):72–78. doi: 10.1128/iai.43.1.72-78.1984

Molecular cloning of the temperature-inducible outer membrane protein 1 of Yersinia pseudotuberculosis.

I Bölin, H Wolf-Watz
PMCID: PMC263390  PMID: 6317574

Abstract

When Yersinia pseudotuberculosis is shifted from growth at 26 degrees C to growth at 37 degrees C, the synthesis of a plasmid-associated outer membrane protein, protein 1, is induced (Bölin et al., Infect. Immun. 37:506-512). The structural gene of this protein was found to be located on the virulence plasmid pIB1 of Y. pseudotuberculosis. One cosmid hybrid plasmid pBW8 was studied which carried a region of the virulence plasmid. This hybrid plasmid expressed in Escherichia coli K-12 a novel temperature-inducible outer membrane protein which is immunologically related to and has the same molecular weight as protein 1. Protein 1 was purified to homogeneity, and 14 amino acids of the N-terminal end were determined. From this sequence, the tentative corresponding DNA sequence was deduced, and a set of 11-nucleotide-long DNA probes was chemically synthesized. By using these probes in Southern blotting experiments, the genetic location of the N-terminal end of protein 1 was established. By introducing the transposon Tn5 into the virulence plasmid pIB1, mutants were obtained that did not express protein 1. One class of these mutants was still Ca2+ dependent and virulent, suggesting that protein 1 is not a major virulence determinant. Tn5-derived insertion mutants were also obtained which were Ca2+ independent. Such mutants were found to be avirulent. One Ca2+-independent mutant still expressed protein 1, indicating that the regulatory expression of protein 1 is not linked to Ca2+ dependence.

Full text

PDF
72

Images in this article

73Image
on p.73
74Image
on p.74
74Image
on p.74
75Image
on p.75
76Image
on p.76

Selected References

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

  1. 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]
  2. BRUBAKER R. R., SURGALLA M. J. Genotypic alterations associated with avirulence in streptomycin-resistant Pasteurella pestis. J Bacteriol. 1962 Oct;84:615–624. doi: 10.1128/jb.84.4.615-624.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. BURROWS T. W., BACON G. A. V and W antigens in strains of Pasteurella pseudotuberculosis. Br J Exp Pathol. 1960 Feb;41:38–44. [PMC free article] [PubMed] [Google Scholar]
  5. BURROWS T. W. VIRULENCE OF PASTEURELLA PESTIS AND IMMUNITY TO PLAGUE. Ergeb Mikrobiol Immunitatsforsch Exp Ther. 1963;37:59–113. doi: 10.1007/978-3-662-36742-1_2. [DOI] [PubMed] [Google Scholar]
  6. 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. doi: 10.1016/0147-619x(81)90019-6. [DOI] [PubMed] [Google Scholar]
  7. Bochner B. R., Huang H. C., Schieven G. L., Ames B. N. Positive selection for loss of tetracycline resistance. J Bacteriol. 1980 Aug;143(2):926–933. doi: 10.1128/jb.143.2.926-933.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Brubaker R. R. The genus Yersinia: biochemistry and genetics of virulence. Curr Top Microbiol Immunol. 1972;57:111–158. doi: 10.1007/978-3-642-65297-4_4. [DOI] [PubMed] [Google Scholar]
  9. Bölin I., Norlander L., Wolf-Watz H. Temperature-inducible outer membrane protein of Yersinia pseudotuberculosis and Yersinia enterocolitica is associated with the virulence plasmid. Infect Immun. 1982 Aug;37(2):506–512. doi: 10.1128/iai.37.2.506-512.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Carter P. B., Zahorchak R. J., Brubaker R. R. Plague virulence antigens from Yersinia enterocolitica. Infect Immun. 1980 May;28(2):638–640. doi: 10.1128/iai.28.2.638-640.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Elmblad A., Josephson S., Palm G. Synthesis of mixed oligodeoxyribonucleotides following the solid phase method. Nucleic Acids Res. 1982 May 25;10(10):3291–3301. doi: 10.1093/nar/10.10.3291. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Eriksson U., Sundelin J., Rask L., Peterson P. A. The NH2-terminal amino acid sequence of cellular retinoic-acid binding protein from rat testis. FEBS Lett. 1981 Nov 30;135(1):70–72. doi: 10.1016/0014-5793(81)80945-3. [DOI] [PubMed] [Google Scholar]
  13. Ferber D. M., Brubaker R. R. Plasmids in Yersinia pestis. Infect Immun. 1981 Feb;31(2):839–841. doi: 10.1128/iai.31.2.839-841.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gemski P., Lazere J. R., Casey T. Plasmid associated with pathogenicity and calcium dependency of Yersinia enterocolitica. Infect Immun. 1980 Feb;27(2):682–685. doi: 10.1128/iai.27.2.682-685.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gemski P., Lazere J. R., Casey T., Wohlhieter J. A. Presence of a virulence-associated plasmid in Yersinia pseudotuberculosis. Infect Immun. 1980 Jun;28(3):1044–1047. doi: 10.1128/iai.28.3.1044-1047.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. HIGUCHI K., SMITH J. L. 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. doi: 10.1128/jb.81.4.605-608.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hager D. A., Burgess R. R. Elution of proteins from sodium dodecyl sulfate-polyacrylamide gels, removal of sodium dodecyl sulfate, and renaturation of enzymatic activity: results with sigma subunit of Escherichia coli RNA polymerase, wheat germ DNA topoisomerase, and other enzymes. Anal Biochem. 1980 Nov 15;109(1):76–86. doi: 10.1016/0003-2697(80)90013-5. [DOI] [PubMed] [Google Scholar]
  18. Hohn B., Collins J. A small cosmid for efficient cloning of large DNA fragments. Gene. 1980 Nov;11(3-4):291–298. doi: 10.1016/0378-1119(80)90069-4. [DOI] [PubMed] [Google Scholar]
  19. LAWTON W. D., ERDMAN R. L., SURGALLA M. J. BIOSYNTHESIS AND PURIFICATION OF V AND W ANTIGEN IN PASTEURELLA PESTIS. J Immunol. 1963 Aug;91:179–184. doi: 10.21236/ad0299868. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Perry R. D., Brubaker R. R. Vwa+ phenotype of Yersinia enterocolitica. Infect Immun. 1983 Apr;40(1):166–171. doi: 10.1128/iai.40.1.166-171.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Portnoy D. A., Blank H. F., Kingsbury D. T., Falkow S. Genetic analysis of essential plasmid determinants of pathogenicity in Yersinia pestis. J Infect Dis. 1983 Aug;148(2):297–304. doi: 10.1093/infdis/148.2.297. [DOI] [PubMed] [Google Scholar]
  23. Portnoy D. A., Falkow S. Virulence-associated plasmids from Yersinia enterocolitica and Yersinia pestis. J Bacteriol. 1981 Dec;148(3):877–883. doi: 10.1128/jb.148.3.877-883.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. 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]
  26. Suggs S. V., Wallace R. B., Hirose T., Kawashima E. H., Itakura K. Use of synthetic oligonucleotides as hybridization probes: isolation of cloned cDNA sequences for human beta 2-microglobulin. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6613–6617. doi: 10.1073/pnas.78.11.6613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Swanson J., Mayer L. W., Tam M. R. Antigenicity of Neisseria gonorrhoeae outer membrane protein(s) III detected by immunoprecipitation and Western blot transfer with a monoclonal antibody. Infect Immun. 1982 Nov;38(2):668–672. doi: 10.1128/iai.38.2.668-672.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Trägårdh L., Curman B., Wiman K., Rask L., Peterson P. A. Chemical, physical-chemical, and immunological properties of papain-solubilized human transplatation antigens. Biochemistry. 1979 May 29;18(11):2218–2226. doi: 10.1021/bi00578a013. [DOI] [PubMed] [Google Scholar]
  29. Zink D. L., Feeley J. C., Wells J. G., Vanderzant C., Vickery J. C., Roof W. D., O'Donovan G. A. Plasmid-mediated tissue invasiveness in Yersinia enterocolitica. Nature. 1980 Jan 10;283(5743):224–226. doi: 10.1038/283224a0. [DOI] [PubMed] [Google Scholar]

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

RESOURCES