MOLECULAR GENETIC ANALYSIS OF YEAST TAFS

我们的长期目标是了解 RNA聚合酶II对真核mRNA基因转录的调节 (Pol II）。 全面了解转录调控 如果我们要理解正常的 以及病理过程和条件，两者都是 基因转录事件。 我们的研究利用基因 易处理的面包酵母，酿酒酵母，作为模型系统， 阐明了因子TFIID在复杂的 mRNA基因转录过程。 这些研究是一个 我们正在进行的TBP分析的结果，TATA盒结合蛋白 它与大多数mRNA编码基因的启动子结合， 启动导致转录的事件链， 表情 TBP似乎不是以游离蛋白的形式存在于细胞内， 但仅在与RNA聚合酶特异性 称为TBP相关的其他TBP结合蛋白的集合 因子（TAF）。 在酵母细胞中，至少有11种TAF， 与TBP结合形成TFIID。 从大量不同的工作 研究人员已经证明，多亚基TFIID复合物是 与控制mRNA基因的调节事件密切相关 转录。 因此，了解的组成和功能， TFIID TBP-TAF复合物以及其他不同的TBP-TAF复合物将 是弄清基因转录是如何控制的关键。 的 我们将在实验中使用的方法将是多方面的 结合生化和遗传分析，重点是酵母TBP 我们克隆的编码多种酵母TAF的基因 form. 我们将研究这些因素与TBP的相互作用， 其他，启动子DNA和RNA聚合酶II。 成功完成 这些实验将大大增加我们对宇宙万物的理解。 pol Ⅱ介导的mRNA基因转录的重要过程。

Our long term goal is to understand the molecular mechanisms of regulation of eukaryotic mRNA gene transcription by RNA Polymerase II (Pol II). A comprehensive knowledge of transcriptional regulatory mechanisms is absolutely essential if we are to understand both normal and pathological processes and conditions, both of which are products of gene transcription events. Our studies utilize the genetically tractable Baker's Yeast, Saccharomyces cerevisiae, as a model system for elucidating the detailed role that factor TFIID plays in the complicated process of mRNA gene transcription by Pol II. These studies are an outgrowth of our ongoing analyses of TBP, the TATA-box Binding Protein which binds to the promoter of most mRNA encoding genes in order to set into motion the chain of events leading to transcription and thus gene expression. TBP appears not to exist as a free protein inside the cell, but is only functional when associated with RNA Polymerase-specific collections of additional TBP binding proteins known as TBP Associated Factors (TAFs). In yeast cells there are at least eleven TAFs which associate with TBP to form TFIID. Work from a large number of different investigators has demonstrated that the multisubunit TFIID complex is intimately involved in the regulatory events which control mRNA gene transcription. Hence, understanding the composition and function of the TFIID TBP-TAF complex, as well as other distinct TBP-TAF complexes, will be key to figuring out how gene transcription is controlled. The approach which we will use in our experiments will be multi-faceted combining biochemical and genetic analyses which focus upon yeast TBP and the genes encoding the multiple yeast TAFs which we have in cloned form. We will examine the interplay of these factors with TBP, each other, promoter DNA, and RNA polymerase II. Successful completion of these experiments will greatly increase our understanding of the all important process of mRNA gene transcription mediated by pol II.