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Unusual Neutral Oligosaccharides in Mature Sindbis Virus Glycoproteins are Synthesized from Truncated Precursor Oligosaccharides in Chinese Hamster Ovary Cells

Department of Microbiology & Immunology, University of Louisville School of Medicine, Louisville, Kentucky 40292, U.S.A.

We have previously demonstrated the presence of unusual small asparaginyloligosaccharides [(Man)₃GlcNAc₂-ASN] in the mature glycoproteins of Sindbis virus released from both wild-type and lectin-resistant Chinese hamster ovary cells, but the mechanism of synthesis of these structures was not determined. Gel filtration and endo--N-acetylglucosaminidase analyses of Pronase-digested glycopeptides from [³H]mannose-labelled Sindbis virus released at different times after infection of a phytohaemagglutinin-resistant line of Chinese hamster ovary cells demonstrated that these small asparaginyl-oligosaccharides were present in similar relative amounts in virus released throughout the virus infection, rather than arising primarily at late times when cytopathic effects were maximal. Similar analyses of pulse-labelled, cell-associated viral glycopeptides suggested that these small oligosaccharides on mature virus glycoprotein resulted from the normal 1,2-mannosidase processing of truncated precursor oligosaccharides (containing five rather than nine mannoses), rather than from aberrant processing or degradation of the full-size precursor oligosaccharides or normal intermediates.

Keywords: Sindbis virus, glycoproteins, oligosaccharides

Received 15 July 1982; accepted 23 September 1982.