A role for ATP and TFIIH in activation of the RNA polymerase II preinitiation complex prior to transcription initiation
A role for ATP and TFIIH in activation of the RNA polymerase II preinitiation complex prior to transcription initiation
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DOI：
10.1074/jbc.271.13.7245
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发表时间：
1996-03-29
影响因子：
4.8
通讯作者：
Conaway, RC
中科院分区：
文献类型：
--
作者：
Dvir, A;Garrett, KP;Conaway, RC
在通过RNA聚合酶II合成启动子特异性转录物的前8-10个键中需要ATP辅因子是已经确定的。然而，ATP是否是合成第一个磷酸二酯键所必需的，还是在合成新生转录本的稍晚阶段所必需的，仍然存在争议。古德里奇和Tjian（古德里奇，J.A.，和Tjian，R，（1994）Cell 77，145-156）最近提出通过RNA聚合酶II合成启动子特异性转录物的第一个磷酸二酯键不依赖于ATP和一般转录因子TFIIE和TFIIH。在这里，我们研究了这个模型，总的来说，我们的发现表明ATP、TFIIE和TFIIH可以对转录起始的效率具有深远的影响。首先，我们观察到在用RNA聚合酶II、TFIIH和重组TBP重建的转录系统中，在腺病毒2主要晚期启动子处起始的转录物的第一个磷酸二酯键的合成强烈依赖于ATP、TFIIE和TFIIH，TFIIB、TFIIE和TFIIF。第二，我们证明，在这个酶系统中，ATP依赖的转录起始激活可以在新生转录物的第一个磷酸二酯键合成之前立即发生。最后，我们证明，激活的起始复合物是不稳定的，在抑制剂ATP γ S存在下迅速衰减到失活状态。（腺苷5 '-O-（硫代）三磷酸），甚至在失败转录物的重复合成期间。
A requirement for an ATP cofactor in synthesis of the first 8-10 bonds of promoter-specific transcripts by RNA polymerase II is well established. Whether ATP is required for synthesis of the first phosphodiester bond or at a slightly later stage in synthesis of nascent transcripts, however, remains controversial. Goodrich and Tjian (Goodrich, J. A., and Tjian, R, (1994) Cell 77, 145-156) recently proposed that synthesis of the first phosphodiester bond of promoter-specific transcripts by RNA polymerase II is independent of ATP and general transcription factors TFIIE and TFIIH. Here we investigate this model, Taken together, our findings indicate that ATP, TFIIE, and TFIIH can have a profound effect on the efficiency of transcription initiation, First, we observe that synthesis of the first phosphodiester bond of transcripts initiated at the adenovirus 2 major late promoter depends strongly on ATP, TFIIE, and TFIIH in a transcription system reconstituted with RNA polymerase II, TFIIH, and recombinant TBP, TFIIB, TFIIE, and TFIIF. Second, we demonstrate that, in this enzyme system, ATP-dependent activation of transcription initiation can occur immediately prior to synthesis of the first phosphodiester bond of nascent transcripts, Finally, we demonstrate that the activated initiation complex is unstable and decays rapidly to an inactive state in the presence of the inhibitor ATP gamma S (adenosine 5'-O-(thio)triphosphate), even during reiterative synthesis of abortive transcripts.