STRUCTURAL STUDIES OF Sp1 AND YY1, AND TARGETS IN TFIID

这项拟议的研究项目的主要目标是结合分子生物学和X射线结晶学方法在原子水平上阐明转录激活的机制。首先，我将重点确定人TAF(II)55以及人TAF(II)55-TAF(II)250复合体的X射线结构，该复合体是被认为是许多转录调节蛋白靶标的预启动复合体的一部分。然后，我将进行一系列实验，以定量描述先前报道的已知转录调控因子(如SPL和YY1)的相互作用，并绘制这些蛋白质的结构域，这些蛋白质直接与TAF(II)55-TAF(II)250异二聚体相互作用。为了更详细地描述这些相互作用，我将尝试结晶由这两个TAFs组成的复合体以及转录激活子的相互作用结构域，并解决它们的结构。这些研究将帮助我们了解参与癌症和关节炎的选定的、特征良好的转录激活剂与预启动复合体中的基本因子相互作用激活转录的机制。

The broad aim of the proposed research project is to elucidate the mechanisms of transcriptional activation at the atomic level using a combination of molecular biology and X-ray crystallography approaches. Initially, I will focus on determining the X-ray structure of human TAF(II)55 as well as the human TAF(II)55-TAF(II)250 complex, a part of the pre-initiation complex thought to be a target for a number of transcription regulatory proteins. I will then carry out a series of experiments to quantitatively characterize the previously reported interactions of known regulators of transcription, such as Spl and YY1, and map the domains of these proteins that interact directly with the TAF(II)55-TAF(II)250 heterodimer. In order to characterize these interactions in greater detail, I will attempt to crystallize the complexes composed of both TAFs as well as the interacting domains of the transcriptional activators, and solve their structures. The studies will help us understand the mechanisms by which selected, well-characterized transcriptional activators involved in cancer and arthritis interact with the basal factors in the pre-initiation complex to activate transcription.