MOLECULAR ANALYSIS OF RNAPIII TRANSCRIPTION INITIATION

RNAPIII 转录起始的分子分析

• 批准号: 2685079
• 负责人: Steven M Hahn
• 金额: $ 25.13万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-09-30 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2685079
• 关键词: DNA binding protein; DNA directed RNA polymerase; fungal genetics; genetic transcription; molecular cloning; mutant; polymerase chain reaction; protein structure function; site directed mutagenesis; transcription factor

DESCRIPTION: The broad, long-term objectives of this proposal are to determine at a molecular level the mechanism of transcription initiation by RNA polymerase III and how this relates to the initiation mechanisms for the other nuclear polymerases. This will lead to a better understanding of the transcription initiation machinery and how it is regulated. This is important because the transcription machinery is the ultimate target of many signal transduction pathways and gene-specific regulatory factors. Understanding the fundamental process of gene control will provide a basis for understanding many types of diseases such as cancer, which often result from aberrant regulation of transcription initiation. In addition, other diseases resulting from defects in repair of DNA damage are caused by mutations in components of the transcription machinery. The specific aims of this project are designed to study through molecular, genetic and biochemical methods, the function of TFIIIB, a multi-subunit component of the Pol III transcription machinery. Dr. Hahn will complete the cloning of the TFIIIB subunit TFIIIB90 and perform a molecular and genetic analysis of its function. Dr. Hahn will continue the genetic and biochemical analysis of BRF1, a second subunit of TFIIIB. He will also use a genetic approach to understand the role of TATA binding protein, a third subunit of TFIIIB, in Pol III transcription Finally, Dr. Hahn has devised a strategy to examine in detail the role of the individual TFIIIB subunits in the process of polymerase recruitment and open complex formation. His work will involve molecular techniques (gene cloning, mutagenesis, and PCR), biochemical fractionation, in vitro transcription, protein biochemistry, and yeast molecular genetics.

描述：本提案的广泛、长期目标是 在分子水平上确定转录起始的机制 RNA聚合酶III以及这与起始机制的关系 其他的核聚合酶。这将导致更好的 理解转录起始机制以及它是如何 监管.这一点很重要，因为转录机制是 许多信号转导通路的最终靶点和基因特异性 调节因素。了解基因的基本过程 控制将为了解许多类型的疾病提供基础 例如癌症，其通常是由于 转录起始此外，其他疾病引起的 DNA损伤修复的缺陷是由基因突变引起的。 转录机器 该项目的具体目标旨在研究 分子、遗传和生物化学方法，TFIIIB的功能， Pol III是Pol III转录机制的多亚基组分。博士 Hahn将完成TFIIIB亚基TFIIIB 90的克隆， 对其功能进行分子和遗传分析。哈恩医生会 继续进行BRF1的遗传和生化分析， TFIIIB的。他还将使用遗传学的方法来了解 TATA结合蛋白，TFIIIB的第三个亚基，在Pol III中， 最后，哈恩博士设计了一种策略， 详细说明各个TFIIIB亚单位在治疗过程中的作用。 聚合酶募集和开放复合物形成。他的工作将包括 分子技术（基因克隆、诱变和PCR）、生物化学 分离、体外转录、蛋白质生物化学和酵母 分子遗传学