1887

Abstract

The rickettsial pathogen undergoes a variation in which virulent isolates (phase 1) become avirulent (phase 2) after repeated passage in a non-immunologically competent host. Biochemically, this variation is associated with a lipopolysaccharide modification and possibly other factors. Genetically, the regions of DNA responsible for phase variation have not been identified. We have sought to determine whether the plasmid identified in acute disease isolates, QpHl, which represents approximately 5% of the coding capacity of this organism is involved in phase variation. Plasmids from phase 1 and phase 2 variants (designated QpHl and QpH2, respectively) were compared by restriction endonuclease digestion and Southern blot hybridization to determine whether sequence changes in the phase 2 plasmid might account for changes in the virulence of phase 2 organisms compared with that of phase 1 cells. Using over 20 different restriction enzymes, no changes in DNA restriction fragment patterns were detected regardless of whether the phase change occurred during egg or tissue culture passage. The plasmid-specific mRNAs produced from metabolically active, purified cells were identical for each phase type. Using QpH1 or QpH2 DNA as a template, the mRNA produced by an extract was also identical. Finally, the proteins encoded by either plasmid in an transcription/translation reaction were identical. These data indicate that within the limits of our analysis, the plasmid DNA from phase variants is structurally and functionally the same and is therefore unlikely to be involved in phase variation.

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1988-07-01
2022-01-29
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References

  1. Baca O., Paretsky D. 1983; Q-fever and Coxiella burnetii: a model for host parasite interactions. Microbiological Reviews 47:127–149
    [Google Scholar]
  2. Baca O., Martinez I. L., Aragon A. S., Klassen D. 1980; Isolation and partial characterization of lipopolysaccharide from phase II Coxiella burnetii. Canadian Journal of Microbiology 26:819–825
    [Google Scholar]
  3. Bobb C., Downs C. M. 1962; The phase antigens of Coxiella burnetii. Canadian Journal of Microbiology 8:689–701
    [Google Scholar]
  4. Bonner W. M., Laskey R. A. 1974; A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. European Journal of Biochemistry 46:83–88
    [Google Scholar]
  5. Corbin D., Ditta G., Helinski D. R. 1982; Clustering of nitrogen fixation (nif) genes in Rhizobium meliloti. Journal of Bacteriology 149:221–228
    [Google Scholar]
  6. Elwell L. P., Shipley P. L. 1980; Plasmid-mediated factors associated with virulence of bacteria to animals. Annual Review of Microbiology 34:465–496
    [Google Scholar]
  7. Fiset P. 1957; Phase variation of Rickettsia (Coxiella) burnetii - a study of the antibody response in guinea pigs and rabbits. Canadian Journal of Microbiology 2:310–321
    [Google Scholar]
  8. Fiset P., Ormsbee R. A. 1968; The antibody response to antigens of Coxiella burnetii. Zentralblatt für Bakteriologie Parasitenkunde Infektionskrankheiten und Hygiene(Abteilung I, Originate A) 206:321–329
    [Google Scholar]
  9. Hackstadt T., Williams J. C. 1981; Biochemical stratagem for obligate parasitism of eukaryotic cells by Coxiella burnetii. Proceedings of the National Academy of Sciences of the United States of America 78:3240–3244
    [Google Scholar]
  10. Hackstadt T., Peacock M. G., Hitchcock P. J., Cole R. L. 1985; Lipopolysaccharide variation in Coxiella burnetii:intrastrain heterogeneity in structure and antigenicity. Infection and Immunity 48:359–365
    [Google Scholar]
  11. Hendrix L. R., Mallavia L. P. 1984; Active transport of proline by Coxiella burnetii. Journal of General Microbiology 130:2857–2863
    [Google Scholar]
  12. Hoyer G. H., Ormsbee R. A., Fiset P., Lackman D. B. 1963; Differentiation of phase I and phase II Coxiella burnetii by equilibrium density gradient sedimentation. Nature; London: 197:573–574
    [Google Scholar]
  13. Kazar J., Skultetyova E., Brezina R. 1975; Phagocytosis of Coxiella burnetii by macrophages. Acta virologica 19:426–431
    [Google Scholar]
  14. Kishimoto R. A., Walker J. S. 1976; Interaction between Coxiella burnetii and guinea pig peritoneal macrophages. Infection and Immunity 14:416–421
    [Google Scholar]
  15. Krauss H., Scheifer H. G., Schmatz H. D. 1977; Ultrastructural investigations on surface structures involved in Coxiella burnetii phase variation. Infection and Immunity 15:890–896
    [Google Scholar]
  16. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227:680–685
    [Google Scholar]
  17. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular Cloning, A Laboratory Manual Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  18. O’Rourke A. T., Peacock M. G., Samuel J. E., Frazier M. E., Natvig D. O., Mallavia L. P., Baca O. G. 1985; Genomic analysis of phase I and phase II Coxiella burnetii with restriction endonucleases. Journal of General Microbiology 131:1543–1546
    [Google Scholar]
  19. Rigby P. W., Dieckmann M., Rhodes C., Berg P. 1977; Labelling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. Journal of Molecular Biology 113:237–245
    [Google Scholar]
  20. Samuel J. E., Frazier M. E., Kahn M. L., Thomashow L. S., Mallavia L. P. 1983; Isolation and characterization of a plasmid from phase I Coxiella burnetii. Infection and Immunity 41:488–493
    [Google Scholar]
  21. Samuel J. E., Frazier M. E., Mallavia L. P. 1985; Correlation of plasmid type and disease caused by Coxiella burnetii. Infection and Immunity 49:775–779
    [Google Scholar]
  22. Schramek S., Brezina R. 1979; Isolation of an endotoxic lipopolysaccharide from phase II Coxiella burnetii. Acta virologica 23:349
    [Google Scholar]
  23. Schramek S., Mayer H. 1982; Different sugar compositions of lipopolysaccharide isolated from phase I and pure phase II cells of Coxiella burnetii. Infecton and Immunity 38:53–57
    [Google Scholar]
  24. Southern E. M. 1975; Detection of specific sequences among DNA fragments separated by gel electrophoresis. Journal of Molecular Biology 98:503–517
    [Google Scholar]
  25. Stoker M. P. G., Fiset P. 1956; Phase variation of the Nine Mile and other strains of Rickettsia burnetii. Canadian Journal of Microbiology 3:435–445
    [Google Scholar]
  26. Vodkin M. H., Williams J. C., Stephenson E. H. 1986; Genetic heterogeneity among isolates of Coxiella burnetii. Journal of General Microbiology 132:455–463
    [Google Scholar]
  27. Williams J. C., Stewart S. 1984; Identification of immunogenic proteins of Coxiella burnetii phase variants. Microbiology 84:257–262
    [Google Scholar]
  28. Williams J. C., Peacock M. G., Mccaul T. F. 1981; Immunological and biological characterization of Coxiella burnetii, phase I and phase II, separated from host components. Infection and Immunity 32:840–851
    [Google Scholar]
  29. Yang H., Ivashkiva L., Chen H., Zubay G., Cashel M. 1980; Cell-free coupled transcription-translation system for investigation of linear DNA segments. Proceedings of the National Academy of Sciences of the United States of America 77:7029–7033
    [Google Scholar]
  30. Yanisch-Perron C., Vierra J., Messing J. 1985; Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33:103–119
    [Google Scholar]
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