Core Promoter-selective Transcription by RNA Polymerase II

智力上的优点。真核基因的表达在转录水平上由序列特异性DNA结合调节剂调节，所述序列特异性DNA结合调节剂控制“核心启动子”处的通用/基础RNA聚合酶II转录机器的组装和活性。核心启动子包括基因的转录起始位点，并且由各种核心启动子DNA元件组成，包括TATA盒、起始子和在不同基因中不连续存在的其他元件。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异性增强子和转录调节子的选择性应答，因此在差异基因表达的调节中起关键作用。最近的分析揭示了从酵母到人类真核基因核心启动子中Initiator元件的保守性和普遍性。然而，引发剂的功能和TATA-引发剂协同作用的机制仍然知之甚少。 该项目将表征启动子功能和TATA-启动子协同作用的机制，以及转录因子和最近鉴定的辅因子在体外和体内启动子依赖性基因转录中的作用。将在体外重建的转录反应中进行机制分析，并将通过多种方法分析体内基因选择性转录功能。 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，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。将通过研究出版物和科学会议的讨论向科学界传播和提供成果和生成的工具，并通过互联网向更广泛的受众传播和提供。PI致力于增加在科学中代表性不足的少数民族的参与，并将继续通过在滨江的加州大学建立少数民族访问计划从事研究少数民族学生。

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.