Novel glycosylation pathways of retroviral envelope proteins identified with avian reticuloendotheliosis virus.
Previously, we identified two mature glycoproteins, gp90, the surface glycoprotein, and gp20, the transmembrane protein, from avian reticuloendotheliosis virus and an avian reticuloendotheliosis virus env gene-encoded intracellular polyprotein gPr77env, but the precise relationship of gPr77env to the mature envelope proteins was not determined (W.-P. Tsai, T.D. Copeland, and S. Oroszlan, Virology 155:567-583, 1986). In the present study, using metabolic labeling of viral proteins with [35S]cysteine, radioimmunoprecipitation, and carbohydrate structure analysis, we have identified a higher-molecular-weight endo-H-resistant env gene-encoded polyprotein designated gPr115env in addition to the endo-H-sensitive gPr77env. It appears that gPr77env is the primary polyprotein precursor, modified with mannosyloligosaccharides that are processed into sialic-acid-rich extraordinarily large complex-type carbohydrates (up to 17 kilodaltons for each N-linked site) on the gp90 domain but not on the gPr22 domain. In this process, gPr77env is converted into the apparently endo-H-resistant secondary polyprotein, gPr115env, which is rapidly processed into gp90 and gPr22. The proteolytic processing which occurs only after the appearance of an endo-H resistant precursor is now clearly demonstrated for a retrovirus. Some important aspects of carbohydrate structure, including the site-specific glycosylation, as well as the intracellular location and nature of the potential enzyme involved in the proteolytic cleavage of gPr115env are discussed.
Tsai
W P
WP
Laboratory of Molecular Virology and Carcinogenesis, NCI-Frederick Cancer Research Facility, Maryland 21701.
Oroszlan
S
S
D000818:Animals; D001345:Autoradiography; D002241:Carbohydrates / Q000032:analysis; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000032:analysis; D006026:Glycoside Hydrolases / Q000378:metabolism; D006031:Glycosylation; D007118:Immunoassay; D011498:Protein Precursors / Q000032:analysis; D001356:Reticuloendotheliosis virus / Q000378:metabolism*; D012190:Retroviridae / Q000378:metabolism*; D012191:Retroviridae Proteins / Q000378:metabolism*; D014759:Viral Envelope Proteins / Q000378:metabolism*
D002241:Carbohydrates; D006023:Glycoproteins; D011498:Protein Precursors; D012191:Retroviridae Proteins; D014759:Viral Envelope Proteins; D006026:Glycoside Hydrolases