Minvielle-Sebastia L, Preker P J, Keller W
Department of Cell Biology, University of Basel, Switzerland.
Science. 1994 Dec 9;266(5191):1702-5. doi: 10.1126/science.7992054.
Most eukaryotic pre-messenger RNAs are processed at their 3' ends by endonucleolytic cleavage and polyadenylation. In yeast, this processing requires polyadenylate [poly(A)] polymerase (PAP) and other proteins that have not yet been characterized. Here, mutations in the PAP1 gene were shown to be synergistically lethal with previously identified mutations in the RNA14 and RNA15 genes, which suggests that their encoded proteins participate in 3'-end processing. Indeed, extracts from ma14 and rna15 mutants were shown to be deficient in both steps of processing. Biochemical complementation experiments and reconstitution of both activities with partially purified cleavage factor I (CF I) validated the genetic prediction.
大多数真核生物前体信使RNA在其3'末端通过核酸内切酶切割和聚腺苷酸化进行加工。在酵母中,这种加工需要聚腺苷酸[poly(A)]聚合酶(PAP)和其他尚未被鉴定的蛋白质。在这里,PAP1基因突变被证明与先前鉴定的RNA14和RNA15基因突变具有协同致死性,这表明它们编码的蛋白质参与3'末端加工。事实上,ma14和rna15突变体的提取物在加工的两个步骤中都表现出缺陷。生化互补实验以及用部分纯化的切割因子I(CF I)对两种活性的重建验证了遗传预测。