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转录激活和抑制机制都可能涉及TFIID。

Mechanisms of transcriptional activation and repression can both involve TFIID.

作者信息

Manley J L, Um M, Li C, Ashali H

机构信息

Department of Biological Sciences, Columbia University, New York, NY 10027, USA.

出版信息

Philos Trans R Soc Lond B Biol Sci. 1996 Apr 29;351(1339):517-26. doi: 10.1098/rstb.1996.0050.

Abstract

Regulation of transcription involves the activities of activators and repressors. Recent experiments have provided evidence that the function of both types of regulators can involve interactions with one or more component of the basal transcription machinery. A principal target appears to be TFIID, which consists of the TATA binding protein (TBP) and associated factors (TAFs). Here we describe experiments that provide added support for the idea that interactions affecting TFIID can play important roles in both activation and repression. We show, using transfection assays in Drosophila Schneider cells, that recruitment of TBP to a promoter as a GAL4-TBP fusion protein can provide a substantial activation of transcription. The conserved core of TBP is necessary and sufficient for this effect, which was observed with both TATA-containing and TATA-lacking basal promoters. These findings extend experiments performed in yeast, and strengthen the idea that recruitment of TBP (TFIID) can be an important mechanism of activation. We also provide further support for the idea that TBP can be a target for a transcriptional repressor, the Drosophila Even-skipped protein (Eve). We present evidence that the homeodomain, which is necessary for binding TBP in vitro, can also be required for repression in vivo, independent of its role in DNA binding. On the other hand, deletion of the alanine/proline-rich region that is essential for repression in vivo and TBP binding in vitro does not significantly affect DNA binding by the purified protein. These results strengthen the view that TBP, either directly or indirectly as a component of TFIID, can be a target of both activators and repressors.

摘要

转录调控涉及激活因子和阻遏因子的活性。最近的实验提供了证据,表明这两种调控因子的功能都可能涉及与基础转录机制的一个或多个组分的相互作用。一个主要靶点似乎是TFIID,它由TATA结合蛋白(TBP)和相关因子(TAFs)组成。在这里,我们描述了一些实验,这些实验为以下观点提供了更多支持:影响TFIID的相互作用在激活和阻遏过程中都能发挥重要作用。我们利用果蝇Schneider细胞中的转染实验表明,作为GAL4-TBP融合蛋白将TBP招募到启动子上可以显著激活转录。TBP的保守核心对于这种效应是必需且充分的,在含TATA和不含TATA的基础启动子上均观察到了这种效应。这些发现扩展了在酵母中进行的实验,并强化了TBP(TFIID)招募可能是激活的重要机制这一观点。我们还为TBP可以成为转录阻遏因子果蝇Even-skipped蛋白(Eve)的靶点这一观点提供了进一步支持。我们提供的证据表明,在体外结合TBP所必需的同源结构域,在体内阻遏过程中也可能是必需的,这与其在DNA结合中的作用无关。另一方面,删除在体内阻遏和体外TBP结合中必不可少的富含丙氨酸/脯氨酸的区域,对纯化蛋白的DNA结合没有显著影响。这些结果强化了这样一种观点,即TBP作为TFIID的一个组分,无论是直接还是间接,都可以成为激活因子和阻遏因子的靶点。

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