Control of Transcription in Eukaryotic Cells

越来越明显的是，RNA聚合酶II从起始到转录延长的转变是一个复杂的过程。 真核基因表达的主要调控检查点。在目前的支持期间，我们的实验室 在描述这种转变方面取得了重大进展。我们设计了一个工作模型， 启动子清除是这个过程的第一步。我们的清除模式包括一个新的概念， 转录起始因子的重要作用。我们还证明了一个明显的后清除 转换发生在转录起始点下游25-35个碱基处。我们分析了RNA 聚合酶II通过后清除阶段提出了新的见解机制， 这种聚合酶可以在转录的这一点上被捕获。在本申请中，我们提出四个 具体目标，通过这些目标，我们将（i）测试我们的排雷和排雷后过渡初步模型 (ii)确定在这些转换中单个转录因子的作用，和（iii）识别潜在的DMA 对这些转变有重要影响的序列。 与公共卫生的相关性：许多基因的正确表达对人类健康至关重要， 通过转录机器在转录后的一个点上的延长暂停来控制 基因信息的传播已经开始。目前尚不清楚这些“管制闸门”如何运作。为了 为了最终操纵这些门以造福人类健康，必须揭示这些门的分子结构。 涉及的机制。这里提出的研究旨在提供这方面的知识。的间隙 和后清除转换，我们研究的是试管模型的监管点中观察到的 cell.

It is increasingly evident that the transition from initiation to transcript elongation by RNA polymerase II is a major regulatory checkpoint in eukaryotic gene expression. During the current support period our laboratory has made significant advances in characterizing this transition. We have devised a working model for promoter clearance, the initial step in this process. Our clearance model includes the novel concept of an important role for transcript initiation factors. We have also demonstrated that a distinct postclearance transition occurs from 25-35 bases downstream of transcription start. Our analysis of the ability of RNA polymerase II to pass through the postclearance stage has suggested new insights into mechanisms through which polymerase can be trapped at this point in transcription. In the current application, we propose four specific aims, through which we will (i) test our initial models for the clearance and postclearance transitions (ii) determine the roles of individual transcription factors in these transitions, and (iii)identify underlying DMA sequences which have important effects on these transitions. Relevance to Public Health: Many genes whose correct expression is essential for human health are controlled through the extended pausing of the transcriptional machinery at a point just after the transcription of genetic information has initiated. It is not currently known how these "regulated gates" operate. In order to ultimately manipulate these gates for the benefit of human health, it is essential to uncover the molecular mechanisms involved. The research proposed here is aimed at providing this knowledge. The clearance and postclearance transitions which we study are test tube models for the regulatory points observed in the cell.