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首页> 外文期刊>Molecular and Cellular Biology >Genetic analysis of the repetitive carboxyl-terminal domain of the largest subunit of mouse RNA polymerase II.
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Genetic analysis of the repetitive carboxyl-terminal domain of the largest subunit of mouse RNA polymerase II.

机译：小鼠RNA聚合酶II最大亚基的重复羧基末端结构域的遗传分析。

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The carboxyl-terminal domain (CTD) of the mouse RNA polymerase II largest subunit consists of 52 repeats of a seven-amino-acid block with the consensus sequence Tyr-Ser-Pro-Thr-Ser-Pro-Ser. A genetic approach was used to determine whether the CTD plays an essential role in RNA polymerase function. Deletion, insertion, and substitution mutations were created in the repetitive region of an alpha-amanitin-resistant largest-subunit gene. The effects of these mutations on RNA polymerase II activity were assayed by measuring the ability of mutant genes to confer alpha-amanitin resistance after transfection of susceptible rodent cells. Mutations that resulted in CTDs containing between 36 and 78 repeats had no effect on the transfer of alpha-amanitin resistance, whereas mutations with 25 or fewer repeats were inactive in this assay. Mutations that contained 29, 31, or 32 repeats had an intermediate effect; the number of alpha-amanitin-resistant colonies was lower and the colonies obtained were smaller, indicating that the mutant RNA polymerase II was defective. In addition, not all of the heptameric repeats were functionally equivalent in that repeats that diverged in up to three amino acids from the consensus sequence could not substitute for the conserved heptamer repeats. We concluded that the CTD is essential for RNA polymerase II activity, since substantial mutations in this region result in loss of function.
机译：小鼠RNA聚合酶II最大亚基的羧基末端结构域（CTD）由一个具有七个共有序列Tyr-Ser-Pro-Thr-Ser-Pro-Ser的七个氨基酸嵌段组成的52个重复组成。遗传方法用于确定CTD是否在RNA聚合酶功能中起重要作用。删除，插入和替换突变是在一个抵抗α-阿马尼汀的最大亚基基因的重复区域中创建的。这些突变对RNA聚合酶II活性的影响通过在易感啮齿动物细胞转染后测量突变基因赋予α-amanitin抗性的能力进行了分析。导致CTD包含36至78个重复序列的突变对α-amanitin抗性的转移没有影响，而具有25个或更少重复序列的突变在此分析中无效。包含29、31或32个重复的突变具有中间作用;耐α-阿马尼汀的菌落数较少，获得的菌落较小，表明突变体RNA聚合酶II存在缺陷。此外，并非所有的七聚体重复序列在功能上都是等效的，因为从共有序列中最多分裂出三个氨基酸的重复序列不能替代保守的七聚体重复序列。我们得出结论，CTD对于RNA聚合酶II活性至关重要，因为该区域的实质性突变会导致功能丧失。

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