title: Analysis of rubella virus E1 glycosylation mutants expressed in COS cells.
abstract: cDNA clones encoding the envelope glycoprotein E1 of rubella virus (RV) were altered by site-directed mutagenesis at consensus sites for addition of N-linked glycans. The resulting plasmids were introduced into COS cells and the mutant E1 proteins were analyzed by indirect immunofluorescence, radioimmunoprecipitation, and immunoblotting. We found that RV E1 contains three N-linked oligosaccharides, each approximately 2 kDa in size. Although lack of glycosylation did not appear to affect targeting of E1 to the Golgi region, mutants lacking N-linked glycans at Asn 177 and Asn 209 failed to bind anti-E1 antibodies under nonreducing conditions. Our results suggest that glycosylation may be important for expression of important immunologic epitopes on RV E1.
authors.affiliation: Department of Pathology, University of British Columbia Research Center, Vancouver, Canada.
authors.forename: T C
authors.initials: TC
authors.lastname: Hobman
authors.forename: Z Y
authors.initials: ZY
authors.lastname: Qiu
authors.forename: H
authors.initials: H
authors.lastname: Chaye
authors.forename: S
authors.initials: S
authors.lastname: Gillam
mesh_terms: D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D004279:DNA, Viral / Q000096:biosynthesis / Q000737:chemistry; D015870:Gene Expression; D006031:Glycosylation; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D012412:Rubella virus / Q000235:genetics* / Q000648:ultrastructure; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D004279:DNA, Viral; D014759:Viral Envelope Proteins; C061826:E1 envelope protein, Rubella virus