A surface epitope undergoing high-frequency phase variation is shared by Mycoplasma gallisepticum and Mycoplasma bovis.
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.
Yogev
D
D
Department of Membrane and Ultrastructure Research, Hebrew University-Hadassah Medical School, Jerusalem.
Menaker
D
D
Strutzberg
K
K
Levisohn
S
S
Kirchhoff
H
H
Hinz
K H
KH
Rosengarten
R
R
D000818:Animals; D000940:Antigenic Variation; D000942:Antigens, Bacterial / Q000737:chemistry / Q000276:immunology*; D000954:Antigens, Surface / Q000737:chemistry / Q000276:immunology*; D001426:Bacterial Proteins / Q000737:chemistry / Q000276:immunology; D002645:Chickens / Q000276:immunology; D000939:Epitopes; D005798:Genes, Bacterial; D008074:Lipoproteins / Q000276:immunology; D008565:Membrane Proteins / Q000737:chemistry / Q000276:immunology; D008970:Molecular Weight; D009174:Mycoplasma / Q000276:immunology*; D012150:Polymorphism, Restriction Fragment Length
D000942:Antigens, Bacterial; D000954:Antigens, Surface; D001426:Bacterial Proteins; D000939:Epitopes; D008074:Lipoproteins; D008565:Membrane Proteins