Transfer of carbohydrates on to nascent glycoprotein of vesicular stomatitis virus.
I studied the glycosylation in vivo of a viral envelope protein, the glycoprotein of vesicular stomatitis virus (VSV), by pulse labelling of virus-infected HeLa cells with 3H-labelled monosaccharides (mannose, glucosamine). Radioactivity was incorporated into the fraction of membrane-bound polyribosomes, although metabolic conversion of [3H]-mannose into amino acids was negligible. Dissociation of bound polyribosomes revealed that the radioactively co-purified with the peptidyl-tRNA. The nascent peptides were released by alkaline hydrolysis, immunoprecipitated and analysed by polyacrylamide-gel electrophoresis. It is apparent from the size distribution of the [3H]mannose-labelled nascent chains that attachment of carbohydrate starts when approximately half of the amino acid sequence of the viral glycoprotein has been synthesized.
D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D005944:Glucosamine / Q000378:metabolism*; D006023:Glycoproteins / Q000096:biosynthesis*; D006367:HeLa Cells / Q000378:metabolism; D008358:Mannose / Q000378:metabolism*; D011132:Polyribosomes / Q000378:metabolism; D012343:RNA, Transfer / Q000378:metabolism; D014721:Vesicular stomatitis Indiana virus / Q000378:metabolism*; D014764:Viral Proteins / Q000096:biosynthesis*
D006023:Glycoproteins; D014764:Viral Proteins; D012343:RNA, Transfer; D005944:Glucosamine; D008358:Mannose