Deoxyribonucleic acid-dependent ribonucleic acid polymerases from normal and polyoma-transformed BHK-21/C13 cells.
DNA-dependent RNA polymerase (EC 2.7.7.6) ACTIVITIES FROM NORMAL BHK-21/C13 cells and from BHK-21/C13 cells transformed by polyoma virus (PYY cells) were solubilized and fractionated on columns of DEAE-Sephadex. Various properties of the A and B enzymes from the two types of cell were compared. 1. The yields of polymerase relative to the DNA content of the nuclear preparations are similar for both cell types. 2. The ionic-strength optima of polymerases A and B are 12.5 mM and 100mM with respect to (NH4)2SO4 for both cell types. 3. The Mn2+/Mg2+ activity ratio (measured at the respective optimum for each cation) for polymerase A from BHK-21/C13 cells was 1.48 and for the polymerase A from PYY cells was 0.55. The corresponding ratios for polymerase B were 10.11 for BHK-21/C13 cells and 22.75 for PYY cells. 4. Minor differences in the ability of the A polymerases to transcribe native and denatured DNA templates were observed; such differences were not apparent when the B polymerases were compared. 5. All the polymerases were inhibited completely by actinomycin D and by rifampicin AF/013, but not markedly so by rifampicin. Alpha-amanitin inhibited polymerase B but not polymerase A.
D000546:Amanitins / Q000494:pharmacology; D000645:Ammonium Sulfate; D000818:Animals; D002460:Cell Line; D002471:Cell Transformation, Neoplastic; D006224:Cricetinae; D004247:DNA; D012321:DNA-Directed RNA Polymerases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D003609:Dactinomycin / Q000494:pharmacology; D007668:Kidney; D008274:Magnesium; D008345:Manganese; D011120:Polyomavirus; D012293:Rifampin / Q000494:pharmacology; D013698:Templates, Genetic
D000546:Amanitins; D003609:Dactinomycin; D008345:Manganese; D004247:DNA; D012321:DNA-Directed RNA Polymerases; D008274:Magnesium; D000645:Ammonium Sulfate; D012293:Rifampin