Altered synthesis and processing of oligosaccharides of vesicular stomatitis virus glycoprotein in different lectin-resistant Chinese hamster ovary cell lines.
To determine the particular intracellular steps in the glycosylation of the vesicular stomatitis virus (VSV) glycoprotein that were altered in several lectin-resistant CHO cell lines, VSV-infected parental and mutant cells were pulse-labeled for 30 and 120 min with [3H]mannose and [3H]glucosamine. Cell-associated viral glycopeptides were analyzed by gel filtration combined with specific glycosidase digestions and compared with the corresponding mature virion oligosaccharides. The intracellular glycosylation of the VSV glycoprotein in a mutant cell line resistant to phytohemagglutinin was identical to that in the normal cells except for a complete block in processing at a specific step in the final trimming of the oligomannosyl core from five to three mannoses. The results demonstrated that a double-mutant cell line selected from the phytohemagglutinin-resistant cells for resistance to concanavalin A had an additional defect in one of the earliest stages of glycosylation, resulting in smaller precursor oligosaccharides linked to protein.
D000818:Animals; D002460:Cell Line; D002850:Chromatography, Gel; D003208:Concanavalin A / Q000494:pharmacology; D006224:Cricetinae; D004351:Drug Resistance; D005260:Female; D006023:Glycoproteins / Q000096:biosynthesis*; D009154:Mutation; D009844:Oligosaccharides / Q000096:biosynthesis*; D010053:Ovary; D010835:Phytohemagglutinins / Q000494:pharmacology; D014721:Vesicular stomatitis Indiana virus / Q000378:metabolism*; D014764:Viral Proteins / Q000096:biosynthesis*
D006023:Glycoproteins; D009844:Oligosaccharides; D010835:Phytohemagglutinins; D014764:Viral Proteins; D003208:Concanavalin A