Core Promoter-selective Transcription by RNA Polymerase II

智慧的功绩。真核基因的表达在转录水平上受到序列特异性DNA结合调控因子的调控，这些调控因子控制着“核心启动子”处一般/基础RNA聚合酶II转录机制的组装和活性。核心启动子包含基因的转录起始点，由各种核心启动子DNA元件组成，包括TATA盒、启动子和其他在不同基因中不同存在的元件。这些DNA元件不仅控制核心启动子的强度，而且控制核心启动子对特定增强子和转录调节因子的选择性反应，因此在基因差异表达的调控中起着关键作用。最近的分析揭示了真核基因核心启动子中的启动子元件的保守性和普遍性，从酵母到人类。然而，引发剂的功能和TATA-引发剂协同作用的机理仍然知之甚少。本项目将对启动子功能和TATA-启动子协同作用的机制以及转录因子和最近发现的辅因子在启动子依赖的基因体外和体内转录中的作用进行表征。机制分析将在体外重组的转录反应中进行，体内的基因选择性转录功能将通过多种方法进行分析。这些结果有望改变我们对真核生物基因表达调控中最基本的过程之一的理解。更广泛的影响。该项目将通过继续指导本科生和研究生以及指导博士后研究人员来加强研究和培训的基础设施。这项研究融入了PI对哺乳动物基因调控的长期兴趣，并将与他的教学职责协同工作。成果和生成的工具将通过研究出版物和科学会议上的讨论向科学界传播和提供，并通过互联网向更广泛的受众提供。国际学生联合会致力于增加少数族裔对科学的参与，并将继续通过加州大学河滨分校已建立的少数族裔准入计划，让少数族裔学生参与研究。

Intellectulal Merit. Expression of eukaryotic genes is regulated at the transcriptional level by sequence-specific DNA-binding regulators that control the assembly and activity of the general/basal RNA polymerase II transcription machinery at the "core promoter". The core promoter encompasses the transcription initiation site of the gene and is composed of various core promoter DNA elements including the TATA box, the Initiator and other elements that are variably present in different genes. These DNA elements control not only the strength of the core promoter but also the selective response of the core promoter to specific enhancers and transcription regulators and therefore play a critical role in the regulation of differential gene expression. Recent analyses have revealed the conservation and prevalence of the Initiator element in eukaryotic gene core promoters from yeast to humans. However, the functions of the Initiator and the mechanisms of TATA-Initiator synergy, have remained poorly understood. This project will characterize the mechanisms of Initiator function and TATA-Initiator synergy and the roles of transcription factors and recently identified cofactors in Initiator-dependent gene transcription in vitro and in vivo. Mechanistic analyses will be performed in reconstituted transcription reactions in vitro, and gene-selective transcription functions in vivo will be analyzed by a number of approaches. The results promise to transform our understanding of the one of the most fundamental processes involved in regulation of gene expression in eukaryotes. Broader Impacts. This project will enhance the infrastructure of research and training through the continued instruction of undergraduate and graduate students and mentoring of postdoctoral researchers. The research integrates into the PI's long-term interest in mammalian gene regulation and will be synergistic with his teaching responsibilities. Results and generated tools will be communicated and made available to the scientific community via research publications and through discussions in scientific meetings, and to broader audiences via the internet. The PI is committed to increase the participation of underrepresented minorities in science and will continue to engage minority students in research through established minority access programs at the University of California at Riverside.