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首页> 外文期刊>Molecular and Cellular Biology >The Two Steps of Poly(A)-Dependent Termination, Pausing and Release, Can Be Uncoupled by Truncation of the RNA Polymerase II Carboxyl-Terminal Repeat Domain
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The Two Steps of Poly(A)-Dependent Termination, Pausing and Release, Can Be Uncoupled by Truncation of the RNA Polymerase II Carboxyl-Terminal Repeat Domain

机译：可通过截断RNA聚合酶II羧基末端重复结构域来解开依赖于Poly（A）的终止，暂停和释放的两个步骤

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摘要

The carboxyl-terminal repeat domain (CTD) of RNA polymerase II is thought to help coordinate events during RNA metabolism. The mammalian CTD consists of 52 imperfectly repeated heptads followed by 10 additional residues at the C terminus. The CTD is required for cleavage and polyadenylation in vitro. We studied poly(A)-dependent termination in vivo using CTD truncation mutants. Poly(A)-dependent termination occurs in two steps, pause and release. We found that the CTD is required for release, the first 25 heptads being sufficient. Neither the final 10 amino acids nor the variant heptads of the second half of the CTD were required. No part of the CTD was required for poly(A)-dependent pausing—the poly(A) signal could communicate directly with the body of the polymerase. By removing the CTD, pausing could be observed without being obscured by release. Poly(A)-dependent pausing appeared to operate by slowing down the polymerase, such as by down-regulation of a positive elongation factor. Although the first 25 heptads supported undiminished poly(A)-dependent termination, they did not efficiently support events near the promoter involved in abortive elongation. However, the second half of the CTD, including the final 10 amino acids, was sufficient for these functions.
机译：RNA聚合酶II的羧基末端重复域（CTD）被认为有助于协调RNA代谢过程中的事件。哺乳动物CTD由52个不完全重复的七足动物组成，然后在C末端有10个其他残基。体外裂解和聚腺苷酸化需要CTD。我们研究了体内使用CTD截断突变体的poly（A）依赖终止。取决于Poly（A）的终止发生在两个步骤中，即暂停和释放。我们发现，释放CTD是必需的，前25个七足体就足够了。 CTD的后半部分不需要最后的10个氨基酸或变体七肽。依赖poly（A）的暂停不需要CTD的任何部分，因为poly（A）信号可以直接与聚合酶体内进行通讯。通过移除CTD，可以观察到暂停，而不会被释放遮盖。依赖于Poly（A）的暂停似乎是通过减慢聚合酶的速度来进行的，例如通过下调正伸长因子来实现。尽管最初的25个七足动物支持不减弱的poly（A）依赖性终止，但它们并不能有效地支持启动子附近参与流产延伸的事件。但是，CTD的后半部分（包括最后的10个氨基酸）足以实现这些功能。

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