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Stepwise Recruitment of Components of the Preinitiation Complex by Upstream Activators In Vivo

机译：体内上游活化剂逐步招募预起始复合物的组分

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Recently, it was found that if either the TATA binding protein or RNA polymerase II holoenzyme is artificially tethered to a promoter, transcription is activated. This finding provided presumptive evidence that upstream activating proteins function by recruiting components of the preinitiation complex (PIC) to the promoter. To date, however, there have been no studies demonstrating that upstream factors actually recruit components of the PIC to the promoter in vivo. Therefore, we have studied the mechanism of action of two disparate transactivating domains. We present a series of in vivo functional assays that demonstrate that each of these proteins targets different components of the PIC for recruitment. We show that, by targeting different components of the PIC for recruitment, these activating domains can cooperate with each other to activate transcription synergistically and that, even within one protein, two different activating subdomains can activate transcription synergistically by cooperating to recruit different components of the PIC. Finally, considering our work together with previous studies, we propose that certain transcription factors both recruit components of the PIC and facilitate steps in transcriptional activation that occur subsequent to recruitment.
机译：最近，发现如果将TATA结合蛋白或RNA聚合酶II全酶人工连接至启动子，则转录被激活。该发现提供了推测性的证据，即上游激活蛋白通过将预起始复合物（PIC）的成分募集至启动子而发挥功能。然而，迄今为止，还没有研究表明上游因素实际上在体内将PIC的成分募集到启动子上。因此，我们研究了两个不同的反式激活域的作用机理。我们提出了一系列的体内功能测定法，证明这些蛋白质中的每一个都靶向PIC的不同成分进行募集。我们表明，通过靶向PIC的不同成分进行募集，这些激活域可以相互协作以协同激活转录，并且即使在一种蛋白质内，两个不同的激活亚域也可以通过协同募集不同的PIC成分来协同激活转录。 PIC。最后，考虑到我们的工作和以前的研究，我们建议某些转录因子既募集PIC的组成部分，又促进募集后发生的转录激活步骤。

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