In vitro synthesis of Rous sarcoma virus-specific RNA is catalyzed by a DNA-dependent RNA polymerase.
Synthesis of Rous sarcoma virus RNA was examined in vitro with a new assay for radioactive virus-specific RNA. Nuclei from infected and uninfected cells were incubated with ribonucleoside [alpha-(32)P]triphosphates, Mn(++), Mg(++) and (NH(4))(2)SO(4). Incorporation into total and viral RNA proceeded with similar kinetics for up to 25 min at 37 degrees . About 0.5% of the RNA synthesized by the infected system was scored as virus-specific, compared to 0.03% of the RNA from the uninfected system and 0.005% of the RNA synthesized by monkey kidney cell nuclei. Preincubation with DNase or actinomycin D completely suppressed total and virus-specific RNA synthesis. alpha-Amanitin, a specific inhibitor of eukaryotic RNA polymerase II, completely inhibited virus-specific RNA synthesis, while reducing total RNA synthesis by only 50%. We conclude that tumor virus-specific RNA is synthesized on a DNA template, most probably by the host's RNA polymerase II.
D000645:Ammonium Sulfate; D000818:Animals; D001358:Avian Sarcoma Viruses / Q000378:metabolism*; D001487:Basidiomycota; D002467:Cell Nucleus / Q000378:metabolism; D002474:Cell-Free System; D002478:Cells, Cultured; D002645:Chickens; D004247:DNA; D012321:DNA-Directed RNA Polymerases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D003609:Dactinomycin / Q000494:pharmacology; D003851:Deoxyribonucleases / Q000494:pharmacology; D003864:Depression, Chemical; D005347:Fibroblasts; D007700:Kinetics; D008274:Magnesium; D008345:Manganese; D009693:Nucleic Acid Hybridization; D009842:Oligopeptides / Q000494:pharmacology; D010456:Peptides, Cyclic / Q000494:pharmacology; D010761:Phosphorus Radioisotopes; D012367:RNA, Viral / Q000096:biosynthesis* / Q000302:isolation & purification; D012265:Ribonucleotides; D013698:Templates, Genetic
D009842:Oligopeptides; D010456:Peptides, Cyclic; D010761:Phosphorus Radioisotopes; D012367:RNA, Viral; D012265:Ribonucleotides; D003609:Dactinomycin; D008345:Manganese; D004247:DNA; D012321:DNA-Directed RNA Polymerases; D003851:Deoxyribonucleases; D008274:Magnesium; D000645:Ammonium Sulfate