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• W2004085162 abstract "SUMMARY The initial env gene polyproteins present in Moloney murine leukaemia virus (M-MuLV)-infected NIH/3T3 cells were examined to determine their relationship to each other as well as their role in generating env gene products gp70, p15(E) and p12(E). Steady-state labelling with [3H]glucosamine revealed anti-gp69/71 immunoprecipitable proteins of mol. wt. 93000 (gPr93env), 83000 (gPr83env) and 70000 (gp70), whereas similar labelling with [3H]fucose showed only two bands of anti-gp69/71 immunoprecipitable radioactivity migrating in SDS-polyacrylamide gels with gPr93env and gp70. Pulse-chase experiments employing [3H]leucine labelling instead of labelled sugars failed to detect gPr93env using similar techniques. The gPr83env was the only polypeptide detected in 15 min [3H]leucine pulselabellings, whereas gPr83env, gp70, p15(E) and p12(E) were detected in chase experiments using appropriate antisera in immunoprecipitation experiments. Pretreatment of infected cells with tunicamycin, an inhibitor of glycosylation, allowed the synthesis of a major band at mol. wt. 62000 (Pr62env) and a minor band of 73000 mol. wt. at the expense of gPr83env. In pulse-chase experiments conducted in the presence of tunicamycin, Pr62env increased during the early chase period but disappeared during the later stages of the chase. No product of Pr62env was detected. Cation-exchange chromatography of tryptic digests of radioactive tyrosine-labelled gPr83env, Pr62env and gp70 showed sequence relationships among the three proteins. Comparison of the two-dimensional fingerprints of [3H]leucine-labelled gPr83env and the mature proteins gp70 and p12(E) support their precursor-product relationship. Of interest is the observation that gp70 and p12(E) seemed to share a few leucine-containing tryptic peptides. These results provide strong evidence that gPr83env is the primary product of the env gene which, upon tunicamycin treatment, is synthesized as a subglycosylated protein, Pr62env. It appears that gPr83env undergoes further modification of its core oligosaccharide structure as detected by fucosylation to yield gPr93env. Our inability to detect gPr93env by [3H]leucine labellings suggests a close chronological relationship between fucosylation and cleavage of the precursor polyprotein, suggesting that cleavage of gPr93env yields gp70 and p15(E). The latter is further cleaved to yield p12(E) plus a polypeptide containing the C-terminal end of p15(E)." @default.
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• W2004085162 date "1982-04-01" @default.
• W2004085162 modified "2023-09-25" @default.
• W2004085162 title "Processing of the env Gene Products of Moloney Murine Leukaemia Virus" @default.
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• W2004085162 doi "https://doi.org/10.1099/0022-1317-59-2-329" @default.
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