Fan Xiaochun, Shi Hua, Adelman Karen, Lis John T
Department of Molecular Biology and Genetics, Cornell University, Biotechnology Building, Ithaca, NY 14853, USA.
Proc Natl Acad Sci U S A. 2004 May 4;101(18):6934-9. doi: 10.1073/pnas.0401523101. Epub 2004 Apr 21.
The TATA-binding protein (TBP) is a critical general transcription factor that associates with the core promoter and acts as a nexus for gene regulation through its interactions with other factors. A large number of proteins recognize the relatively small yet highly conserved C-terminal domain of TBP. One subset of these proteins (general transcription factors) interacts with the TBP.TATA complex and RNA polymerase II to create the preinitiation complex. To study TBP functions in preinitiation complex and other complexes, we generated a set of RNA aptamers with high affinity to yeast TBP. These aptamers act on TBP in different ways: all of them bind TBP competitively with DNA bearing the TATA element, and some can actively disrupt the TBP.TATA interaction in preformed, higher-order complexes containing the additional general transcription factors TFIIB and TFIIA. In crude cell extracts, the aptamers inhibit transcription in ways that reveal the dynamic nature of TBP interactions during initiation and reinitiation.
TATA 结合蛋白(TBP)是一种关键的通用转录因子,它与核心启动子结合,并通过与其他因子的相互作用作为基因调控的枢纽。大量蛋白质识别 TBP 相对较小但高度保守的 C 末端结构域。这些蛋白质的一个子集(通用转录因子)与 TBP-TATA 复合物和 RNA 聚合酶 II 相互作用以形成前起始复合物。为了研究 TBP 在起始前复合物和其他复合物中的功能,我们生成了一组对酵母 TBP 具有高亲和力的 RNA 适体。这些适体以不同方式作用于 TBP:它们都与带有 TATA 元件的 DNA 竞争性结合 TBP,并且一些可以在含有额外通用转录因子 TFIIB 和 TFIIA 的预先形成的高阶复合物中主动破坏 TBP-TATA 相互作用。在粗细胞提取物中,适体以揭示起始和重新起始过程中 TBP 相互作用动态性质的方式抑制转录。