TAFS AND GENERAL TRANSCRIPTION FACTORS IN YEAST

描述(摘自摘要)：启动第II类基因表达 在真核生物中涉及形成一个大的多亚单位复合体 含有RNA聚合酶II和启动子上的附加因子。 塔塔结合蛋白(TBP)在这一过程中起着核心作用，因为它与 直接连接到启动子DNA。至少五种不同的TBP相关蛋白 已发现在第II类基因中有作用的复合体 转录，但它们对基因表达调控的贡献 这些TBP结合蛋白包括TAFII、NC2 (DR1-DRAP1)、MOT1、NOTS和SPTS。遗传学和生物化学实验 在本提案中描述的是旨在调查性质和 TAFIIs以外的TBP结合蛋白在控制第II类分子中的作用 基因在酿酒酵母中的表达。这项建议的四个具体目标 1)研究RNA聚合酶II之间的遗传相互作用 体内全酶和TBP结合蛋白，2)研究其作用 TBP结合蛋白突变对第II类基因表达的影响 Vivo，3)纯化和鉴定各种亚基的组成 TBP结合蛋白参与酿酒酵母中II类基因的表达 使用生化方法，4)研究转录功能 利用野生型和突变型蛋白获得TBP结合蛋白 体外转录系统。

DESCRIPTION (adapted from abstract): Initiation of class II gene expression in eukaryotes involves the formation of a large multisubunit complex containing RNA polymerase II and additional factors on promoters. TATA-binding protein (TBP) plays a central role in this process as it binds directly to promoter DNA. At least five different TBP-associated protein complexes have been identified that have roles in class II gene transcription, but their contributions to the regulation of gene expression are not yet well understood These TBP-binding proteins include TAFIIs, NC2 (Dr1-DRAP1), MOT1, NOTs and SPTs. The genetic and biochemical experiments described in this proposal are designed to investigate the nature and function of TBP-binding proteins other than TAFIIs in control of class II gene expression in S. cerevisiae. The four specific aims of this proposal are: 1) to study the genetic interaction betwee RNA polymerase II holoenzyme and TBP-binding proteins in vivo, 2) to investigate the effects of mutations in the TBP-binding proteins on class II gene expression in vivo, 3) to purify and identify the subunit composition of the various TBP-binding proteins involved in class II gene expression in S. cerevisiae using biochemical approaches, 4) to study the transcriptional functions of the TBP-binding proteins using wild-type and mutant proteins in transcription systems in vitro.