Mechanisms of eukaryotic transcription activation

真核转录激活机制

• 批准号: 8049212
• 负责人: Steven M Hahn
• 金额: $ 60.43万
• 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/8049212
• 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(申请人提供)：转录激活是许多信号转导和发育途径的最终终点，而了解激活机制是了解基因调控的关键。以前的研究表明，通过基因特异性转录激活子和辅助激活子的突变扰乱正常的基因调控可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剂和辅助激活剂用来调节RNA聚合酶II转录的机制。这项拟议的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。这项工作的具体目的是利用生化、结构和分子遗传学方法来检测两个激活域和两个共激活子的直接靶标。我们将研究几个酸性激活剂-辅激活剂复合体的结构，以了解激活剂如何具体识别他们的目标，激活剂-靶识别的共同原理，以及更广泛的，内在无序蛋白质的功能。我们将研究SAGA共激活子与TBP(TATA结合蛋白)的相互作用，以及这种相互作用是如何通过乙酰化来调节的。使用一组独特的交联剂，我们将检查转录机制中介体共激活物复合体的直接靶标。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。综合考虑，我们的结果将导致一个分子模型，说明激活剂如何识别他们的辅助激活剂靶标，以及辅激活剂如何通过与转录机制的直接相互作用来刺激基因表达。 公共卫生相关性：项目叙述本研究的目的是了解转录的机制和调节，即信使核糖核酸的合成过程。转录调控是控制细胞生长、分化和发育的关键步骤之一，而转录缺陷直接导致许多人类疾病。了解转录及其调控的机制将为理解导致多种癌症以及心脏病、神经疾病和出生缺陷的转录紊乱的分子缺陷奠定基础。