Polylactosaminoglycan modification of a small integral membrane glycoprotein, influenza B virus NB.
The structure of the carbohydrate components of NB, the small integral membrane glycoprotein of influenza B virus, was investigated. The carbohydrate chains of NB are processed from the high-mannose form (NB18) to a heterogeneous form of much higher molecular weight, designated NBp. Selection of this carbohydrate-containing form of NB with Datura stramonium lectin, its susceptibility to digestion by endo-beta-galactosidase, and determination of the size of NBp glycopeptides by gel filtration chromatography suggested that the increase in molecular weight is due to processing to polylactosaminoglycan. Investigation of the polypeptides produced by influenza B/Lee/40 virus infection of several cell types and another strain of influenza B virus suggested that the signal for modification to polylactosaminoglycan is contained in NB. Expression of mutants of NB lacking either one or both of the normal N-terminal sites of asparagine-linked glycosylation indicated that both carbohydrate chains are modified to contain polylactosaminoglycan. NBp and a small amount of unprocessed NB18 are expressed at the infected-cell surface, as determined by digestion of the surfaces of intact cells with various endoglycosidases. Unglycosylated NB, expressed either in influenza B virus-infected cells treated with tunicamycin or in cells expressing the NB mutant lacking both N-linked glycosylation sites, was expressed at the cell surface, indicating that NB does not require carbohydrate addition for transport.
Williams
M A
MA
Department of Biochemistry, Molecular Biology, and Cell Biology, Northwestern University, Evanston, Illinois 60208.
Lamb
R A
RA
D000606:Amino Sugars / Q000032:analysis* / Q000378:metabolism; D000818:Animals; D002236:Carbohydrate Conformation; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects; D002850:Chromatography, Gel; D006023:Glycoproteins / Q000032:analysis* / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D007118:Immunoassay; D009981:Influenza B virus / Q000187:drug effects* / Q000648:ultrastructure; D008565:Membrane Proteins / Q000032:analysis; D008970:Molecular Weight; D011134:Polysaccharides / Q000032:analysis* / Q000378:metabolism; D014764:Viral Proteins / Q000032:analysis* / Q000378:metabolism
D000606:Amino Sugars; D006023:Glycoproteins; D008565:Membrane Proteins; C051094:NB glycoprotein, Influenza B virus; D011134:Polysaccharides; D014764:Viral Proteins; C039448:lactosaminoglycan