MOLECULAR GENETIC ANALYSES OF YEAST TAFS

我们的长期目标是了解调控的分子机制 真核生物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过程和条件。 我们的研究利用面包酵母，酿酒酵母，作为模型 系统，用于阐明因子TFIID在 RNA聚合酶II的mRNA基因转录的复杂过程。 这些 研究是我们最近关于TATA盒结合的TBP工作的产物 蛋白质，我们目前知道它是至少两种不同RNA的亚基 聚合酶II特异性多蛋白复合物：（1）它包含DNA- TFIID的结合亚基和（2）含有至少一个TFIID结合亚基的复合物， 另外的蛋白质称为TAF 170，TBP相关的Mr因子， 170,000. 大量不同调查人员的工作 证明了多亚基，含TBP的TFIID复合物是 与实际控制mRNA的调节事件密切相关 基因转录 因此，了解的组成和功能， TFIID/TBP和其他含TBP的复合物是了解基因如何表达的关键。 转录是可控的。 我们将使用的方法是我们的 实验将是多方面的结合生化和遗传 分析集中在酵母TBP和编码多种酵母TBP的基因上， 我们最近克隆的TAF。 我们将研究这些因素之间的相互作用 因子与TBP、彼此、启动子DNA和RNA聚合酶II。 这些实验的成功完成将大大提高我们的 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.