Mann C, Buhler J M, Treich I, Sentenac A
Cell. 1987 Feb 27;48(4):627-37. doi: 10.1016/0092-8674(87)90241-8.
Yeast RNA polymerases A and C share an approximately equal to 40 kd subunit. We have identified, sequenced, and mutagenized in vitro the AC40 subunit gene. The RPC40 gene is unique in the yeast genome and is required for cell viability. This gene contains an open reading frame encoding a 37.6 kd protein having no significant homology with bacterial RNA polymerase subunits. The promoter region contains a 19 bp sequence also present in the largest subunit of RNA polymerase C. It also contains a well-conserved RPG box, a sequence found in the promoter region of many genes encoding the translational apparatus. A novel, plasmid-shuffling method was developed to isolate a large number of RPC40 ts mutants. One of these, ts4, was shown to be defective in the synthesis of RNA polymerases A and C at the restrictive temperature. In contrast, RNA polymerase B was made normally.
酵母RNA聚合酶A和C共有一个约40kd的亚基。我们已在体外对AC40亚基基因进行了鉴定、测序和诱变。RPC40基因在酵母基因组中是独一无二的,是细胞生存能力所必需的。该基因包含一个开放阅读框,编码一个37.6kd的蛋白质,该蛋白质与细菌RNA聚合酶亚基没有明显的同源性。启动子区域包含一个19bp的序列,该序列也存在于RNA聚合酶C的最大亚基中。它还包含一个保守性良好的RPG框,这是在许多编码翻译装置的基因的启动子区域中发现的序列。我们开发了一种新的质粒洗牌方法来分离大量的RPC40温度敏感型突变体。其中一个突变体ts4在限制温度下被证明在RNA聚合酶A和C的合成中存在缺陷。相比之下,RNA聚合酶B的合成正常。