Partial purification and characterization of DNA-dependent RNA polymerases I and II from cherry salmon (Oncorhynchus masou).
1. DNA-dependent RNA polymerases I and II were purified approx 3900- and 13,000-fold, respectively, from sonicated nuclear extract of cherry salmon (Oncorhynchus masou) liver by DEAE-Sephadex, heparin-Sepharose and DNA-cellulose column chromatography. 2. The purified RNA polymerases exhibited a requirement for four kinds of ribonucleoside 5'-triphosphates, an exogeneous template and divalent cation. 3. The activities of RNA polymerases I and II were inhibited by Actinomycin D (24 micrograms/ml) but not by Rifampicin (200 micrograms/ml). 4. RNA polymerase I preferred native DNA as template, while polymerase II preferred single-stranded DNA. 5. RNA polymerase II was inhibited by a low concentration of alpha-amanitin (0.02 micrograms/ml). RNA polymerase I was also inhibited by the relatively high concentration of alpha-amanitin (IC50 = 100 micrograms/ml and IC70 = 750 micrograms/ml). 6. RNA polymerases from cherry salmon exhibited a higher activity at low temperature than from rat liver.
D000546:Amanitins / Q000494:pharmacology; D000645:Ammonium Sulfate / Q000494:pharmacology; D000818:Animals; D012321:DNA-Directed RNA Polymerases / Q000302:isolation & purification*; D008099:Liver / Q000201:enzymology; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D012318:RNA Polymerase I / Q000032:analysis / Q000302:isolation & purification*; D012319:RNA Polymerase II / Q000032:analysis / Q000302:isolation & purification*; D012474:Salmon; D013696:Temperature
D000546:Amanitins; D008345:Manganese; D012319:RNA Polymerase II; D012321:DNA-Directed RNA Polymerases; D012318:RNA Polymerase I; D008274:Magnesium; D000645:Ammonium Sulfate