MOLECULAR ANALYSIS OF RNAPIII TRANSCRIPTION INITIATION

RNAPIII 转录起始的分子分析

• 批准号: 2192809
• 负责人: Steven M Hahn
• 金额: $ 12.43万
• 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/2192809
• 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：这项提议的广泛、长期目标是 在分子水平上确定转录启动的机制 RNA聚合酶III及其与启动机制的关系 对于其他的核聚合酶。这将导致一个更好的 了解转录启动机制及其原理 受监管的。这一点很重要，因为转录机制是 许多信号转导途径和基因特异性的最终靶点 监管因素。理解基因的基本过程 控制将为了解许多类型的疾病提供基础 例如癌症，这通常是由于对 转录启动。此外，其他由以下原因引起的疾病 DNA损伤修复的缺陷是由DNA损伤的成分突变引起的 转录机器。 这个项目的具体目标是通过研究 分子、遗传和生化方法，TFIIIB的功能，a Pol III转录机制的多亚基组成。Dr。 哈恩将完成TFIIIB亚基TFIIIB90的克隆和 对其功能进行分子和遗传分析。哈恩博士会 继续第二个亚基BRF1的遗传和生化分析 TFIIIB的。他还将使用遗传学方法来理解这一角色 TFIIIB的第三亚单位TATA结合蛋白在Pol III中的表达 最后，哈恩博士设计了一种策略来研究 详细说明各个TFIIIB亚基在过程中的作用 聚合酶募集和开放复合体的形成。他的工作将包括 分子技术(基因克隆、突变和聚合酶链式反应)、生化 分级、体外转录、蛋白质生物化学和酵母 分子遗传学。