Assembly and processing of the disulfide-linked varicella-zoster virus glycoprotein gpII(140).
Varicella-zoster virus (VZV) specifies the synthesis of at least four families of glycoproteins, which have been designated gpI, gpII, gpIII, and gpIV. In this report we describe the assembly and processing of VZV gpII, a structural protein of an apparent Mr of 140,000, which is the homolog of gB of herpes simplex virus. For these studies, we used two anti-gpII monoclonal antibodies which exhibited both complement-independent neutralization activity and inhibition of virus-induced cell-to-cell fusion. Pulse-chase labeling experiments identified a 124,000-Mr intermediate which was chased to the mature 140,000-Mr product when analyzed in nonreducing gels; in the presence of a reducing agent, the native gp140 was cleaved into two closely migrating species (gp66 and gp68). The biosynthesis of VZV gpII was further analyzed in the presence of the following inhibitors of glycoprotein processing: tunicamycin, monensin, castanospermine, swainsonine, and deoxymannojirimycin. All intermediate and mature forms were digested with endoglycosidases H and F, neuraminidase, and O-glycanase to further define high-mannose, complex, and O-linked glycans. Finally, the addition of sulfate residues was investigated. This characterization of VZV gpII revealed the following results. (i) gp128 and gp124 were early high-mannose forms, (ii) gp126 was an intermediate form with complex N-linked oligosaccharides, (iii) gp130 was a later intermediate with both N-linked and O-linked glycans, and (iv) the mature product gp140 contained a mixture of N-linked and O-linked glycans which were both sialated and sulfated. Further investigations indicated that gpII sulfation was inhibited by tunicamycin and castanospermine but not by deoxymannojirimycin or swainsonine. We also concluded that VZV gpII displayed many biological and biochemical properties similar to those of its herpes simplex virus homolog gB.
D017485:1-Deoxynojirimycin; D000118:Acetylglucosaminidase / Q000494:pharmacology; D000470:Alkaloids / Q000494:pharmacology; D000911:Antibodies, Monoclonal / Q000276:immunology; D004220:Disulfides; D005944:Glucosamine / Q000031:analogs & derivatives / Q000494:pharmacology; D006023:Glycoproteins / Q000032:analysis / Q000378:metabolism*; D014645:Herpesvirus 3, Human / Q000276:immunology / Q000378:metabolism* / Q000472:pathogenicity; D006596:Hexosaminidases / Q000494:pharmacology; D006801:Humans; D007212:Indolizines; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D008970:Molecular Weight; D009439:Neuraminidase / Q000494:pharmacology; D013431:Sulfates / Q000378:metabolism; D017026:Swainsonine; D014415:Tunicamycin / Q000494:pharmacology; D014764:Viral Proteins / Q000032:analysis / Q000378:metabolism*; D048809:alpha-N-Acetylgalactosaminidase; D054836:Indolizidines
D017485:1-Deoxynojirimycin; D000118:Acetylglucosaminidase; D000470:Alkaloids; D000911:Antibodies, Monoclonal; D004220:Disulfides; D005944:Glucosamine; D006023:Glycoproteins; D006596:Hexosaminidases; D007212:Indolizines; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D009439:Neuraminidase; D013431:Sulfates; D017026:Swainsonine; D014415:Tunicamycin; D014764:Viral Proteins; D048809:alpha-N-Acetylgalactosaminidase; C482722:NAGA protein, human; C037806:castanospermine; D054836:Indolizidines