Misumi H, Oda T
Acta Med Okayama. 1979 Apr;33(2):91-102.
RNA polymerase was extracted from the Schmidt-Ruppin strain of Rous sarcoma virus (SR-RSV)-induced C3H/He mouse ascites sarcoma cells (SR-C3H). RNA polymerase was separated into RNA polymerases I and II by DEAE-Sephadex chromatography. RNA polymerase I was separated into Ia and Ib fractions by phospho-cellulose chromatography. In SR-C3H cells RNA polymerase Ib was the main component of RNA polymerase I. At 0.05--0.1 M ammonium sulphate RNA polymerase I transcribed native DNA most actively, and RNA polymerase II transcribed denatured DNA most actively. Partial digestion of DNA by DNAase I enhanced RNA synthesis by RNA polymerases I and II. At ionic strength over 0.2 M ammonium sulphate, the initiation reaction of RNA polymerases I and II was inhibited. The initiation complexes of RNA polymerases I and II with native DNA were more stable against high salt concentration than with denatured DNA.
从劳氏肉瘤病毒(SR-RSV)诱导的C3H/He小鼠腹水肉瘤细胞(SR-C3H)的施密特-鲁平株中提取RNA聚合酶。通过DEAE-葡聚糖凝胶色谱法将RNA聚合酶分离为RNA聚合酶I和II。通过磷酸纤维素色谱法将RNA聚合酶I分离为Ia和Ib组分。在SR-C3H细胞中,RNA聚合酶Ib是RNA聚合酶I的主要成分。在0.05-0.1M硫酸铵条件下,RNA聚合酶I转录天然DNA最活跃,而RNA聚合酶II转录变性DNA最活跃。DNA酶I对DNA的部分消化增强了RNA聚合酶I和II的RNA合成。在硫酸铵离子强度超过0.2M时,RNA聚合酶I和II的起始反应受到抑制。RNA聚合酶I和II与天然DNA形成的起始复合物比与变性DNA形成的起始复合物对高盐浓度更稳定。