MOLECULAR GENETIC ANALYSES OF YEAST TAFS

我们的长期目标是了解调控的分子机制 真核细胞的mRNA基因转录。全面了解 转录调控机制是绝对必要的，如果我们要 了解正常和病理的生物学过程和条件。 我们的研究以面包酵母、酿酒酵母为模型 用于阐明因子TFIID在 RNA聚合酶II对mRNA基因转录的复杂过程 研究是我们最近关于TBP-TATA盒结合的工作的结果 蛋白质，我们目前知道它是至少两个不同的RNA的一个亚单位 聚合酶II特异的多蛋白复合体：(1)它由DNA- TFIID的结合亚单位和(2)包含至少一个 MR的附加蛋白TAF170，TBP相关因子约 17万。来自大量不同调查人员的工作 证明了含有TBP的多亚基TFIID复合体是 密切参与实际控制信使核糖核酸的调控事件 基因转录。因此，了解的组成和功能 TFIID/TBP和其他含TBP的复合体是了解基因如何 转录是受控制的。我们将使用的方法是我们的 实验将是多方面的，结合了生化和遗传 酵母TBP和多种酵母编码基因的分析 我们最近克隆了TAFs。我们将研究这些因素之间的相互作用 与TBP、相互作用、启动子DNA和RNA聚合酶II有关的因素。 这些实验的成功完成将大大增加我们的 了解RNA聚合酶II转录的关键过程。

Our long term goal is to understand the molecular mechanisms of regulation of eukaryotic mRNA gene transcription. A comprehensive knowledge of transcriptional regulatory mechanisms is absolutely essential if we are to understand normal and pathological biological processes and conditions. Our studies utilize Bakers 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 RNA polymerase II. These studies are an outgrowth of our recent work on TBP the TATA-box Binding Protein, which we currently know is a subunit of at least two distinct RNA Polymerase II-specific multiprotein complexes: (1) it comprises the DNA- binding subunit of TFIID and (2) a complex containing at least one additional protein termed TAF170, TBP Associated Factor of Mr about 170,000. Work from a large number of different investigators has demonstrated that the multisubunit, TBP-containing TFIID complex is intimately involved in the regulatory events which actually control mRNA gene transcription. Hence, understanding the composition and function of TFIID/TBP and other TBP-containing complexes is key to knowing how gene transcription is controlled. The approach which we will be used i our experiments will be multi-faceted combining biochemical and genetic analyses focusing upon yeast TBP and the genes encoding the multiple yeast TAFs which we have recently cloned. 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 key process of RNA polymerase II transcription.