Mechanisms of eukaryotic transcription activation

真核转录激活机制

基本信息

批准号：
8212257
负责人：
Steven M Hahn
金额：
$ 60.51万
依托单位：
FRED HUTCHINSON CANCER RESEARCH CENTER
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-09-01 至 2014-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8212257
关键词：
Acetylation Affinity Binding Biochemical Biochemical Genetics Biological Biological Assay Code Complement Complex Congenital Abnormality Cross-Linking Reagents DNA Polymerase II Defect Development Disease Eukaryota Gene Expression Gene Expression Regulation General Transcription Factors Genes Genetic Genetic Transcription Heart Diseases Human Laboratories Lead Malignant Neoplasms Maps Mediator of activation protein Messenger RNA Methods Modeling Molecular Molecular Genetics Molecular Models Mutation Pathway interactions Play Property Protein Binding Proteins RNA RNA Polymerase II Reagent Regulation Research Role SAGA Signal Transduction Structure System TATA-Box Binding Protein Testing Transcription Coactivator Transcription Process Transcriptional Activation Transcriptional Regulation Work Yeasts base cancer type cell growth insight molecular modeling nervous system disorder protein complex protein protein interaction public health relevance transcription factor TFIIE yeast genetics

项目摘要

DESCRIPTION (provided by applicant): Summary Activation of transcription is the ultimate endpoint for many signal transduction and developmental pathways, and understanding the mechanism of activation is a key to understanding gene regulation. From previous studies, it is clear that disruption of normal gene regulation by mutations in gene- specific transcription activators and coactivators can lead to cancer and other diseases. The broad long- term objectives of this proposal are to determine the mechanisms used by gene-specific activators and coactivators to regulate RNA polymerase II transcription. The proposed work will provide a basis for understanding gene regulation in normal and diseased states at the molecular level. The specific aims of this work utilize biochemical, structural, and molecular genetic methods to examine the direct targets of two activation domains and two coactivators. We will examine the structure of several acidic activator-coactivator complexes to understand how activators specifically recognize their targets, common principals of activator-target recognition and, more generally, the function of intrinsically disordered proteins. We will examine the interaction of the SAGA coactivator with TBP (TATA binding protein) and how this interaction is regulated by acetylation. Using a unique set of crosslinking reagents, we will examine the direct targets of the Mediator coactivator complex within the transcription machinery. In all cases, we will use yeast molecular genetics to test the functional significance of our biochemical results. Combined, our results will lead to a molecular model for how activators recognize their coactivator targets and how coactivators stimulate gene expression by direct interaction with the transcription machinery. PUBLIC HEALTH RELEVANCE: Project Narrative The objective of this research is to understand the mechanism and regulation of transcription, the process of mRNA synthesis. Regulation of transcription is one of the key steps in control of cell growth, differentiation, and development, and defects in transcription directly contribute to many human illnesses. Understanding the mechanism of transcription and its regulation will form the basis for understanding the molecular defects in transcription disorders leading to many types of cancer, as well as heart disease, neurological disorders, and birth defects.

描述（由申请人提供）：摘要转录激活是许多信号转导和发育途径的最终终点，了解激活机制是理解基因调控的关键。从之前的研究来看，很明显，基因特异性转录激活剂和共激活剂突变对正常基因调控的破坏可能导致癌症和其他疾病。该提案的广泛长期目标是确定基因特异性激活剂和共激活剂用于调节 RNA 聚合酶 II 转录的机制。拟议的工作将为在分子水平上理解正常和患病状态下的基因调控提供基础。这项工作的具体目标是利用生物化学、结构和分子遗传学方法来检查两个激活域和两个共激活子的直接靶标。我们将检查几种酸性激活剂-辅激活剂复合物的结构，以了解激活剂如何特异性识别其靶标、激活剂-靶标识别的常见原理，以及更一般地说，本质上无序的蛋白质的功能。我们将研究 SAGA 共激活剂与 TBP（TATA 结合蛋白）的相互作用以及乙酰化如何调节这种相互作用。使用一套独特的交联试剂，我们将检查转录机制中介体共激活剂复合物的直接靶标。在所有情况下，我们都将使用酵母分子遗传学来测试我们的生化结果的功能意义。结合起来，我们的结果将产生一个分子模型，用于解释激活剂如何识别其共激活剂靶标以及共激活剂如何通过与转录机制直接相互作用来刺激基因表达。公共健康相关性：项目叙述本研究的目的是了解转录的机制和调控以及 mRNA 合成的过程。转录调控是控制细胞生长、分化和发育的关键步骤之一，转录缺陷直接导致许多人类疾病。了解转录及其调控机制将为了解导致多种癌症、心脏病、神经系统疾病和出生缺陷的转录障碍中的分子缺陷奠定基础。