Transcription Regulation by NC2

描述（由申请人提供）：我的实验室的长期目标是 制定一个分子机制的理解运作，以调节 II类基因的表达。为此，本提案的总体目标是 继续并扩大目前正在进行的关于基因 保持在一种压抑的状态。这将通过以下方式实现： 所涉及的蛋白质的表征，以及它们的各种蛋白质-DNA和 蛋白质相互作用Ⅱ类病毒特异性转录的研究 基因已经表明，RNA的大部分复杂性和调控 由RNA聚合酶H催化的合成可能存在于起始步骤 - 启动子序列的识别。已经分离出的蛋白质 一起可以重建来自许多I类启动子的特异性转录。 大部分的重点都放在监管因素的隔离上，例如 活化剂。辅激活子、阻遏子和辅阻遏子。在大多数情况下， 编码这些特定调节因子的基因，以及所有的一般调节因子， 已经克隆了转录因子。然而，一个准确的描述 在转录水平上控制基因表达的机制取决于 从生理学角度理解这一复杂的过程 模板转录场现在已经开始重建 使用染色质模板进行转录。我们的研究启动子识别， RNA聚合酶I转录系统的通用转录因子， 包括镇压和反镇压，对世界的发展有着广泛的影响。 RNA合成的一般机制，并将提供基础， 理解特定的转录（演员可以修改转录 染色质中特定基因或一组基因的活性 和/或使用天然启动子。这些研究很可能会产生 II类基因调控机制的基本原理 可以在分子水平上辨别表达。为此，总体而言， 这项建议的目的是继续和扩大对机制的研究， 其中特异性阻遏物复合物NC 2由两个亚基Dri和 Drapi的作用是调节基因表达。我建议研究使用三个 不同的方法。一种方法是直接了解 因子的生化特性。第二种方法是针对 发展对Dr 1和Drap 1在体内作用的理解，使用 简单的生物，酵母，我们可以应用遗传学和研究 Dr 1和Drap 1与其他转录因子的关系。第三 办法的目的是形成一种理解， 因子在多细胞生物体中，因此我描述了在小鼠中的研究。

DESCRIPTION (provided by applicant): The long-term goal of my laboratory is to formulate an understanding of the molecular mechanisms operating to regulate expression of class II genes. To this end, the overall aim of this proposal is to continue and expand studies now in progress on the mechanisms by which genes are maintained in a repressed state. This will be accomplished through the characterization of the proteins involved, and their various protein-DNA and protein-protein interactions. Studies on specific transcription of class II genes have indicated that most of the complexity and regulation of RNA synthesis catalyzed by RNA polymerase H probably resides in the initiation step - the recognition of promoter sequences. Proteins have been isolated that together can reconstitute specific transcription from many class I promoters. Much of the emphasis has been on the isolation of regulatory factors, such as activators. coactivators, repressors and corepressors. In most instances, the genes encoding these particular regulators, as well as all of the general transcription factors have been cloned. However, a description of the exact mechanisms controlling gene expression at the transcriptional level hinges on an understanding of this complex process mechanistically using physiological templates. The transcription field has now moved towards reconstituting transcription using chromatin templates. Our studies on promoter recognition by the general transcription factors of the RNA polymerase I transcription system, including repression and anti-repression, have broad implications for the mechanism of RNA synthesis in general and will provide the basis for understanding how specific transcription (actors can modify the transcriptional activity of a particular gene or a set of genes within the context of chromatin and/or using natural promoters. It is likely that these studies will yield basic principles from which the regulatory mechanisms of class II gene expression can be discerned at the molecular level. To this end, the overall aim of this proposal is to continue and expand studies on the mechanisms by which a specific repressor complex, NC2, composed of two subunits Dri and Drapi, operates to regulate gene expression. I propose studies using three different approaches. One approach is directed towards understanding the biochemical properties of the factor. The second approach is directed towards developing an understanding of the role of Dr1 and Drap1 in vivo, using a simple organism, yeast, with which we can apply genetics and study the relationships between Dr1 and Drap1 and other transcription factors. The third approach is directed towards formulating an understanding of the role of the factor in a multicellular organism and thus I describe studies in the mouse.