Synthesis and processing of glycoprotein gG of herpes simplex virus type 2.
Monoclonal antibody 13 alpha C5-1-A11 immunoprecipitated two major polypeptides of molecular weights 108,000 and 120,000 from extracts of herpes simplex virus type 2-infected BHK-21 cells labeled with [35S]methionine or [3H]glucosamine. In pulse-chase experiments, both labels were chased from the 120,000-molecular-weight peptide (120K peptide) into the 108K molecule. Endoglycosidase H (endo H) reduced the 120K peptide to a 112K peptide but did not affect the 108K peptide. Similar profiles were obtained with monoclonal antibody AP-1 which reacts with a 92K glycoprotein, gG, which maps to the short unique region of the genome. Cross-absorption experiments indicated that both antibodies reacted with the same peptides, suggesting that the 120K peptide is a partially glycosylated high-mannose-type precursor of gG (pgG1). Immunoprecipitation from monensin-treated cells indicated that pgG1(120K) may undergo peptide cleavage to form a 74K high-mannose-type peptide (pgG2) and that this 74K peptide may be further processed into an endo H-resistant 110K to 116K peptide. In the presence of tunicamycin, gG(108K) was replaced by 110K and 105K peptides which were resistant to both endo H and endoglycosidase F. The 105K peptide was the only molecule labeled by [3H]galactose or [3H]glucosamine in the presence of tunicamycin, and none of the peptides were labeled with [3H]mannose, indicating the probable presence of O-linked sugars in the 105K peptide. Our results imply that cotranslational glycosylation of the unglycosylated precursor 110K peptide results in the high-mannose-type pgG1(120K), which probably undergoes peptide cleavage. This putative cleavage product may then mature into gG (108K) by the trimming of sugars and the addition of complex and probably O-linked sugars; the high-mannose-type pgG2(74K) is probably an intermediate peptide formed in this process.
Balachandran
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Hutt-Fletcher
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D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D006224:Cricetinae; D006023:Glycoproteins / Q000276:immunology / Q000378:metabolism*; D006026:Glycoside Hydrolases / Q000494:pharmacology; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D008985:Monensin / Q000494:pharmacology; D018139:Simplexvirus / Q000378:metabolism*; D014415:Tunicamycin / Q000494:pharmacology; D014759:Viral Envelope Proteins; D014764:Viral Proteins / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D006023:Glycoproteins; D014759:Viral Envelope Proteins; D014764:Viral Proteins; C045464:glycoprotein G, herpes simplex virus type 2; D014415:Tunicamycin; D008985:Monensin; D006026:Glycoside Hydrolases; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase