Candidate Products of Avian Myeloblastosis Virus Oncogene Region Produced by in vitro Translation of Genomic RNA

Abstract

Avian myeloblastosis virus (AMV) genomic RNA was translated in vitro to identify the AMV transforming gene (myb) product(s). Full-length RNA yielded, in addition to the expected Pr76^gag, a gag-related product of 92,000 daltons which did not appear to be a gag-oncogene fusion protein. Subgenomic AMV RNA in vitro translation, nonstructural products (specific for AMV) were of 54,000, 49,000 and 34,000 daltons. Based on their sizes and similar peptide maps for the smaller two proteins, it is proposed that they derive from the myb region of AMV.

Detection and Localization of the v-myb(AMV) Gene Products of Avian Myeloblastosis Virus by a Synthetic Peptide Antiserum

“…The 32K protein could be a degradation product of p46 v-myb(AMv). We feel that its size, however, suggests that it might be an internally initiated product as we (Wright et al, 1984) have discussed previously for the p34 v-myb(AMv) in vitro translation product of AMV genomic RNA. From the FA digest data, the p32 is presumed to represent the carboxy twothirds of p46 V-myb(AMv).…”

“…(Hopp & Woods,198l) of the predicted (Klempnauer et al, 1982) v-myb(AMV)-full-length product. (Wright et al, 1984) with 125I-labelled (Crawford & Lane, 1977) synthetic peptide, boosts were performed intraperitoneally (i.p.) every 4 to 6 weeks with alum precipitates (Hudson & Hay, 1976) of unconjugated synthetic peptide (Sen et al, 1983).…”

“…Labelled cells were lysed and fractionated into nuclear and cytoplasmic preparations as detailed (Neiman et al, 1980). The nuclear pellet was suspended in radioimmunoprecipitation (RIPA) buffer and the cytoplasmic supernatant was brought to the same concentration of RIPA buffer, immunoprecipitated and analysed by PAGE, as described (Wright et al, 1984), except that the Staphylococcus aureus was preincubated with macrophage lysate (macrophages being the normal cell counterpart of the myeloblasts; Moscovici, 1975).…”

“…2 was to remove (Amons & Schrier, 1981). Serum diluted l : 10 in phosphatebuffered saline preabsorbed with 150 ~g BSA per reaction (Wright et al, 1984). Lane 1, cytoplasm, normal serum; lane 2, cytoplasm, anti-synthetic peptide serum; lane 3, nucleus, normal serum; lane 4, nucleus, anti-synthetic peptide serum.…”

“…SDS-PAGE analysis of immunoprecipitation of [35S]cysteine-labelled myeloblast nuclear lysates. Lane 1, preimmune serum; lane 2, anti-synthetic peptide serum; lane 3, anti-synthetic peptide serum preincubated (Wright et al, 1984) with 10 gg synthetic peptide. detergents to increase immunoprecipitation with anti-synthetic peptide sera (Mariottini et al, 1983).…”