Mechanism and Regulation of Core Promoter Recognition and Promoter Escape during Transcription by RNA Polymerase II

智力价值：由DNA编码的基因的表达对维持生命至关重要。基因表达的第一步是通过转录的过程制造基因的RNA拷贝。在此过程中，RNA聚合酶II以DNA为模板合成mRNA分子。这项研究的总体目标是详细了解转录反应的因素和机制，以及如何控制它们。这样的理解是重要的，因为转录是在生长、发育和正常细胞代谢过程中调节基因表达的主要控制点。具体而言，本研究将确定并表征人类细胞核中一个因子的功能，该因子可以加速转录的缓慢步骤，从而定义一种新的调节机制。这项研究还将研究转录发生所必需的含有蛋白质和DNA的大型复合体如何在准备转录的基因上组装。这些研究中使用的方法集中在一个完善的人类转录系统上，该系统是在体外由高度纯化的蛋白质组装而成的。实验利用详细的定量分析，专门研究转录的早期步骤和蛋白质在DNA上的组装。这项研究将影响对蛋白质- dna复合物如何在基因上组装的理解，以及RNA合成的早期步骤如何在细胞中被调节。从这个项目中获得的知识有可能刺激复杂生物系统中转录调控的新研究。更广泛的影响：除了上述的研究，该项目将涉及开发一个本科生物化学实验室的实验模块。本模块将为学生提供设计和实施假设驱动实验研究蛋白质- dna相互作用的经验。此外，学生将接触到新的实验技术，从而加强教学实验课程的科学基础设施。为本科实验室开发实验模块强调了将研究与教育相结合以促进教学、培训和学习的重要性。此外，研究计划的实施将提供机会，投入大量的时间来训练研究生在实验设计，数据解释，创造性思维，准备和写作手稿，并给出正式和非正式的口头报告。

Intellectual merit:The expression of genes that are encoded by DNA is essential for sustained life. The first step in gene expression is making an RNA copy of the gene through a process known as transcription. During this process, the enzyme RNA polymerase II, using DNA as a template, synthesizes mRNA molecules. The broad goal of this research is to arrive at a detailed understanding of the factors and the mechanism of the transcription reaction and how they can be controlled. Such an understanding is important because transcription is a principal control point for regulating gene expression during growth, development, and normal cellular metabolism. Specifically, this research will identify and characterize the function of a factor in human nuclei that can accelerate the rate of the slow step in transcription, hence defining a new mechanism of regulation. This research will also investigate how large complexes containing proteins and DNA, which are required for transcription to occur, assemble at genes which are poised to be transcribed. The methods used in these studies center on a well-established human transcription system that is assembled in vitro from highly purified proteins. The experiments utilize detailed quantitative assays specifically developed to study the early steps of transcription and the assembly of proteins on DNA. The research will impact the understanding of how protein-DNA complexes assemble at genes, and how the early steps in RNA synthesis might be regulated in cells. The knowledge derived from this project has the potential to stimulate new studies of transcriptional regulation in complex biological systems.Broader impacts:In addition to the research described above, the project will involve development of an experimental module for an undergraduate biochemistry laboratory. This module will provide students with experience in designing and implementing a hypothesis-driven experiment investigating protein-DNA interactions. Moreover, students will be exposed to new experimental techniques, thereby enhancing the scientific infrastructure of a teaching laboratory course. Developing an experimental module for an undergraduate laboratory underscores the importance of integrating research and education in order to promote teaching, training, and learning. In addition, implementation of the research plan will provide opportunities to devote a significant amount of time to the training of graduate students in experimental design, interpretation of data, creative thought, preparation and writing of manuscripts, and giving formal and informal oral presentations.