Mechanisms of eukaryotic transcription activation

真核转录激活机制

• 批准号: 7880515
• 负责人: Steven M Hahn
• 金额: $ 52.05万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 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/7880515
• 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

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）：概述转录激活是许多信号转导和发育途径的最终终点，理解激活机制是理解基因调控的关键。从以前的研究中可以清楚地看出，基因特异性转录激活因子和辅激活因子的突变破坏正常的基因调控可能导致癌症和其他疾病。这项提议的广泛的长期目标是确定基因特异性激活因子和共激活因子调节RNA聚合酶II转录的机制。这项工作将为在分子水平上理解正常和疾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研究两个激活结构域和两个辅激活因子的直接靶点。我们将研究几种酸性活化剂-辅活化剂复合物的结构，以了解活化剂如何特异性地识别其靶点，活化剂-靶点识别的共同原理，以及更普遍的内在无序蛋白质的功能。我们将研究佐贺辅激活因子与TBP（TATA结合蛋白）的相互作用，以及这种相互作用如何通过乙酰化调节。使用一套独特的交联试剂，我们将研究转录机制内的介体辅激活因子复合物的直接靶标。在所有情况下，我们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，我们的研究结果将导致一个分子模型，激活剂如何识别其辅激活因子的目标，以及辅激活因子如何刺激基因表达的直接相互作用与转录机制。 公共卫生相关性：本研究的目的是了解转录的机制和调控，mRNA的合成过程。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。理解转录及其调控机制将为理解导致许多类型的癌症以及心脏病、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。