Transcription Regulation by NC2

描述(申请人提供)：我的实验室的长期目标是 形成对调控的分子机制的理解 II类基因的表达。为此，这项提案的总体目标是 继续并扩大目前正在进行的关于基因 都保持在受抑制的状态。这将通过 所涉及的蛋白质及其各种蛋白质的表征-DNA和 蛋白质之间的相互作用。II类基因特异性转录的研究 基因已经表明，RNA的大部分复杂性和调控 由RNA聚合酶H催化的合成可能位于起始步骤 -启动子序列的识别。已经分离出的蛋白质 它们一起可以重组许多I类启动子的特定转录。 很大程度上强调了监管因素的隔离，例如 激活剂。辅活化子、抑制子和辅抑制子。在大多数情况下， 编码这些特定调节器的基因，以及所有通用的 转录因子已经被克隆。然而，对准确的 在转录水平上控制基因表达的机制取决于 生理学对这一复杂过程的机械理解 模板。转录能量场现在已经走向重构 使用染色质模板转录。我们对启动子识别的研究 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 RNA合成的一般机制，并将提供基础 了解特定转录的方式(演员可以修改转录 染色质背景下的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 在分子水平上可以辨别其表达情况。为此，总体上 这项提议的目的是通过以下方式继续和扩大对机制的研究 它是由两个亚基dr1和dr1组成的特定阻遏复合体NC2 DRPI的作用是调节基因的表达。我建议使用三种方法进行研究 不同的方法。一种方法是直接理解 因子的生化性质。第二种方法是针对 了解Dr1和DRap1在体内的作用，使用 简单的生物体，酵母，我们可以用它来应用遗传学和研究 DR1和DRAP1与其他转录因子的关系。第三 方法的目的是形成对 多细胞生物体中的因子，因此我描述了在小鼠身上的研究。

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.