Synthesis and processing of the Marek's disease herpesvirus B antigen glycoprotein complex

The Marek's disease herpesvirus B antigen (MDHV-B) complex was previously immunologically identified and molecularly characterized as a set of three glycoproteins designated gp100, gp60, and gp49 on the basis of apparent molecular weight and immunoprecipitation with both polyclonal and monoclonal antibodies. Immunoprecipitation analysis, previously with polyclonal and more recently with monoclonal antibodies, of infected cell lysates labeled with [35S]methionine in the presence of tunicamycin, an inhibitor of N-linked glycosylation, revealed two putative precursor molecules of 88,000 daltons (pr88) and 44,000 daltons (pr44). High-resolution pulse-chase studies revealed that gp100 was a glycosylated intermediate which was processed to yield gp60 and gp49. This cleavage was inhibited by monensin, an inhibitor of glycoprotein processing. Endo-β-N-acetylglucosaminidases F and H (endo-F, endo-H) reduced gp100 to pr88, indicating that the latter is an intermediate in the biosynthetic pathway. These same enzymes reduced gp49, and to a lesser extent gp60, to pr44, suggesting that pr44 is their polypeptide backbone. Significant support for this concept is the fact that the same monoclonal antibody recognized all three molecules, gp60, gp49, and pr44. In the presence of monensin, terminal addition of complex sugars was also prevented, since gp60 was replaced by a slightly faster migrating component which was insensitive to both endo-F and endo-H. Cell-free translation of infected-cell mRNA, followed by immunoprecipitation analysis with either polyclonal or monoclonal antibody, resulted in detection of a putative unglycosylated precursor polypeptide of 44,000 daltons. Since pr88 was not the initial precursor polypeptide of the MDHV-B complex, its existence may have resulted from dimerization of pr44