Effect of p15-associated protease from an avian RNA tumor virus on avian virus-specific polyprotein precursors.
A proteolytic activity is associated with structural protein p15 in avian RNA tumor viruses. Its effect on the known intracellular viral polyprotein precursors obtained by immunoprecipitation was investigated. Cleavage of Pr76gag resulted in the sequential appearance of p15, p27, and p19. The intracellular precursor Pr180gag-pol was also cleaved by p15, whereas the intracellular glycoprotein precursors of avian RNA tumor viruses, Pr92env, remained unaffected by p15 under all conditions tested. The specificities of the antibodies used to precipitate the precursors influenced the pattern of intermediates and cleavage products obtained by p15 treatment. If virus harvested from the the Prague strain of Rous sarcoma virus, subgroup C-transformed cells at 15-min intervals was incubated at 37 degrees C for further maturation, RNA-dependent DNA polymerase activity showed an optimum of DNA synthesis with 70S viral RNA or synthetic template-primers after short incubation periods. The presence of additional p15 during incubation resulted in a shift of the enzyme activity peak toward earlier time points. Virus harvested at 3-h intervals contained significant amounts of Pr180gag-pol and Pr76gag. The addition of p15 resulted in the cleavage of Pr180gag-pol and Pr76gag, but only a few distinct low-molecular-weight polypeptides appeared. Treatment of purified RNA-dependent DNA polymerase with p15 in vitro resulted in a disappearance of the beta subunit and an enrichment of the alpha subunit. In addition, a polypeptide of 32 x 10(3) molecular weight was generated. The cleavage pattern observed differed from the one obtained by trypsin treatment.
D000818:Animals; D001358:Avian Sarcoma Viruses / Q000032:analysis*; D002642:Chick Embryo; D007122:Immunoelectrophoresis; D010447:Peptide Hydrolases / Q000302:isolation & purification; D011498:Protein Precursors / Q000378:metabolism*; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D014764:Viral Proteins / Q000302:isolation & purification / Q000378:metabolism
D011498:Protein Precursors; D014764:Viral Proteins; D012194:RNA-Directed DNA Polymerase; D010447:Peptide Hydrolases