Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1994 Nov;62(11):4962–4968. doi: 10.1128/iai.62.11.4962-4968.1994

A surface epitope undergoing high-frequency phase variation is shared by Mycoplasma gallisepticum and Mycoplasma bovis.

D Yogev 1, D Menaker 1, K Strutzberg 1, S Levisohn 1, H Kirchhoff 1, K H Hinz 1, R Rosengarten 1
PMCID: PMC303213  PMID: 7523302

Abstract

We have recently reported that three distinct size- and phase-variable surface lipoproteins (Vsps) of the bovine pathogen Mycoplasma bovis possess a common epitope recognized by monoclonal antibody 1E5. In the present study, we show that this epitope is also present on a size-variant protein (PvpA) of the avian pathogen Mycoplasma gallisepticum. Application of monoclonal antibody 1E5 in Western immunoblot analysis of Triton X-114 phase-fractionated proteins and in colony immunoblots, as well as in trypsin and carboxypeptidase digestion experiments, has demonstrated that (i) PvpA is an integral membrane protein with a free C terminus, (ii) the shared epitope is surface exposed, and (iii) PvpA is subjected to high-frequency phase variation in expression. By using serum antibodies from M. gallisepticum-infected chickens, we were able to demonstrate the immunogenic nature of PvpA and identify three additional highly immunogenic Triton X-114 phase proteins (p67, p72, and p75) also undergoing high-frequency phase variation spontaneously and independently. Metabolic labeling experiments with [14C]palmitate and [14C]oleate revealed that PvpA, in contrast to p67, p72, and p75, is not lipid modified. Southern blot hybridization with restriction fragments carrying the pvpA gene of M. gallisepticum or the vspA gene of M. bovis against digested genomic DNA of the two Mycoplasma species indicated the absence of genetic relatedness between the pvpA and vspA genes. The apparent complexity of the antigenic variation phenomenon in M. gallisepticum is discussed.

Full text

PDF
4962

Images in this article

4964Image
on p.4964
4964Image
on p.4964
4965Image
on p.4965
4966Image
on p.4966
4966Image
on p.4966

Selected References

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

  1. Baseman J. B., Morrison-Plummer J., Drouillard D., Puleo-Scheppke B., Tryon V. V., Holt S. C. Identification of a 32-kilodalton protein of Mycoplasma pneumoniae associated with hemadsorption. Isr J Med Sci. 1987 May;23(5):474–479. [PubMed] [Google Scholar]
  2. Behrens A., Heller M., Kirchhoff H., Yogev D., Rosengarten R. A family of phase- and size-variant membrane surface lipoprotein antigens (Vsps) of Mycoplasma bovis. Infect Immun. 1994 Nov;62(11):5075–5084. doi: 10.1128/iai.62.11.5075-5084.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Clyde W. A., Jr, Hu P. C. Antigenic determinants of the attachment protein of Mycoplasma pneumoniae shared by other pathogenic Mycoplasma species. Infect Immun. 1986 Feb;51(2):690–692. doi: 10.1128/iai.51.2.690-692.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dallo S. F., Chavoya A., Baseman J. B. Characterization of the gene for a 30-kilodalton adhesion-related protein of Mycoplasma pneumoniae. Infect Immun. 1990 Dec;58(12):4163–4165. doi: 10.1128/iai.58.12.4163-4165.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hu P. C., Clyde W. A., Jr, Collier A. M. Conservation of pathogenic mycoplasma antigens. Isr J Med Sci. 1984 Oct;20(10):916–919. [PubMed] [Google Scholar]
  6. Jasper D. E. Bovine mycoplasmal mastitis. Adv Vet Sci Comp Med. 1981;25:121–157. [PubMed] [Google Scholar]
  7. Jasper D. E. The role of Mycoplasma in bovine mastitis. J Am Vet Med Assoc. 1982 Jul 15;181(2):158–162. [PubMed] [Google Scholar]
  8. Kenny G. E., Cartwright F. D. Immunoblotting for determination of the antigenic specificities of antibodies to the Mycoplasmatales. Isr J Med Sci. 1984 Oct;20(10):908–911. [PubMed] [Google Scholar]
  9. 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]
  10. Markham P. F., Glew M. D., Whithear K. G., Walker I. D. Molecular cloning of a member of the gene family that encodes pMGA, a hemagglutinin of Mycoplasma gallisepticum. Infect Immun. 1993 Mar;61(3):903–909. doi: 10.1128/iai.61.3.903-909.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Mattsson J. G., Gersdorf H., Göbel U. B., Johansson K. E. Detection of Mycoplasma bovis and Mycoplasma agalactiae by oligonucleotide probes complementary to 16S rRNA. Mol Cell Probes. 1991 Feb;5(1):27–35. doi: 10.1016/0890-8508(91)90035-i. [DOI] [PubMed] [Google Scholar]
  12. Morrison-Plummer J., Lazzell A., Baseman J. B. Shared epitopes between Mycoplasma pneumoniae major adhesin protein P1 and a 140-kilodalton protein of Mycoplasma genitalium. Infect Immun. 1987 Jan;55(1):49–56. doi: 10.1128/iai.55.1.49-56.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Morsy M. A., Panangala V. S., van Santen V. L., Bird R. C. Cloning and partial sequence analysis of a Mycoplasma synoviae DNA fragment encoding epitopes shared with the major adhesin P1 protein of Mycoplasma pneumoniae. Avian Dis. 1993 Oct-Dec;37(4):1105–1112. [PubMed] [Google Scholar]
  14. Panangala V. S., Morsy M. A., Gresham M. M., Toivio-Kinnucan M. Antigenic variation of Mycoplasma gallisepticum, as detected by use of monoclonal antibodies. Am J Vet Res. 1992 Jul;53(7):1139–1144. [PubMed] [Google Scholar]
  15. Pollack Y., Stein R., Razin A., Cedar H. Methylation of foreign DNA sequences in eukaryotic cells. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6463–6467. doi: 10.1073/pnas.77.11.6463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Razin S. Molecular biology and genetics of mycoplasmas (Mollicutes). Microbiol Rev. 1985 Dec;49(4):419–455. doi: 10.1128/mr.49.4.419-455.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Rosengarten R., Behrens A., Stetefeld A., Heller M., Ahrens M., Sachse K., Yogev D., Kirchhoff H. Antigen heterogeneity among isolates of Mycoplasma bovis is generated by high-frequency variation of diverse membrane surface proteins. Infect Immun. 1994 Nov;62(11):5066–5074. doi: 10.1128/iai.62.11.5066-5074.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rosengarten R., Wise K. S. Phenotypic switching in mycoplasmas: phase variation of diverse surface lipoproteins. Science. 1990 Jan 19;247(4940):315–318. doi: 10.1126/science.1688663. [DOI] [PubMed] [Google Scholar]
  19. Rosengarten R., Wise K. S. The Vlp system of Mycoplasma hyorhinis: combinatorial expression of distinct size variant lipoproteins generating high-frequency surface antigenic variation. J Bacteriol. 1991 Aug;173(15):4782–4793. doi: 10.1128/jb.173.15.4782-4793.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  21. Theiss P. M., Kim M. F., Wise K. S. Differential protein expression and surface presentation generate high-frequency antigenic variation in Mycoplasma fermentans. Infect Immun. 1993 Dec;61(12):5123–5128. doi: 10.1128/iai.61.12.5123-5128.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Watson H. L., McDaniel L. S., Blalock D. K., Fallon M. T., Cassell G. H. Heterogeneity among strains and a high rate of variation within strains of a major surface antigen of Mycoplasma pulmonis. Infect Immun. 1988 May;56(5):1358–1363. doi: 10.1128/iai.56.5.1358-1363.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Weinstock R., Sweet R., Weiss M., Cedar H., Axel R. Intragenic DNA spacers interrupt the ovalbumin gene. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1299–1303. doi: 10.1073/pnas.75.3.1299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Wise K. S., Kim M. F., Theiss P. M., Lo S. C. A family of strain-variant surface lipoproteins of Mycoplasma fermentans. Infect Immun. 1993 Aug;61(8):3327–3333. doi: 10.1128/iai.61.8.3327-3333.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Yogev D., Levisohn S., Kleven S. H., Halachmi D., Razin S. Ribosomal RNA gene probes to detect intraspecies heterogeneity in Mycoplasma gallisepticum and M. synoviae. Avian Dis. 1988 Apr-Jun;32(2):220–231. [PubMed] [Google Scholar]
  27. Yogev D., Rosengarten R., Watson-McKown R., Wise K. S. Molecular basis of Mycoplasma surface antigenic variation: a novel set of divergent genes undergo spontaneous mutation of periodic coding regions and 5' regulatory sequences. EMBO J. 1991 Dec;10(13):4069–4079. doi: 10.1002/j.1460-2075.1991.tb04983.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES