Lobban P E, Siminovitch L, Ingles C J
Cell. 1976 May;8(1):65-70. doi: 10.1016/0092-8674(76)90186-0.
Amal, an alpha-amanitin-resistant mutant of the Chinese hamster ovary cell line, contains an RNA polymerase activity which elutes from DEAE-Sephadex at a salt concentration characteristic of an RNA polymerase II, but which is not sensitive to alpha-amanitin at levels where the polymerase II of wild-type cells is strongly inhibited. This result suggests that Amal owes its amanitin-resistant phenotype to a mutation affecting one of its genes for RNA polymerase II. To test this hypothesis, we purified the enzyme from Amal and then compared its properties with those of the wild-type enzyme. The mutant enzyme is indeed a polymerase II, and is over 600 times less sensitive to alpha-amanitin and more thermolabile than the wild-type enzyme.
阿马尔是中国仓鼠卵巢细胞系的一种对α-鹅膏蕈碱具有抗性的突变体,它含有一种RNA聚合酶活性,该活性从DEAE-葡聚糖凝胶上洗脱时的盐浓度具有RNA聚合酶II的特征,但在野生型细胞的聚合酶II受到强烈抑制的水平下,它对α-鹅膏蕈碱不敏感。这一结果表明,阿马尔对鹅膏蕈碱的抗性表型归因于影响其RNA聚合酶II基因之一的突变。为了验证这一假设,我们从阿马尔中纯化了该酶,然后将其性质与野生型酶的性质进行了比较。突变酶确实是一种聚合酶II,并且对α-鹅膏蕈碱的敏感性比野生型酶低600多倍,且更不耐热。