Welsh J D, Leibowitz M J, Wickner R B
Nucleic Acids Res. 1980 Jun 11;8(11):2349-63. doi: 10.1093/nar/8.11.2349.
The "killer" plasmid and a larger double-stranded RNA plasmid of yeast exist in intracellular virion particles. Purification of these particles from a diploid killer strain of yeast (grown into stationary growth on ethanol) resulted in co-purification of a DNA-independent RNA polymerase activity. This activity incorporates and requires all four ribonucleoside triphosphates and will not act on deoxyribonucleoside triphosphates. The reaction requires magnesium, is inhibited by sulfhydryl-oxidizing reagents and high concentrations of monovalent cation, but is insensitive to DNase, alpha-amanitin, and actinomycin D. Pyrophosphate inhibits the reaction as does ethidium bromide. Exogenous nucleic acids have no effect on the reaction. The product is mostly single-stranded RNA, some of which is released from the enzymatically active virions.
酵母的“杀手”质粒和一个更大的双链RNA质粒存在于细胞内病毒粒子中。从酵母的二倍体杀手菌株(在乙醇上生长至稳定期)中纯化这些粒子,结果共纯化出一种不依赖DNA的RNA聚合酶活性。该活性掺入并需要所有四种核糖核苷三磷酸,且不会作用于脱氧核糖核苷三磷酸。该反应需要镁,受巯基氧化试剂和高浓度单价阳离子抑制,但对DNase、α-鹅膏蕈碱和放线菌素D不敏感。焦磷酸和溴化乙锭一样会抑制该反应。外源核酸对该反应无影响。产物大多为单链RNA,其中一些从具有酶活性的病毒粒子中释放出来。