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; 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）：转录的摘要激活是许多信号转导和发育途径的最终终点，理解激活的机理是理解基因调节的关键。从以前的研究中可以明显看出，基因转录激活剂和共激活剂中突变对正常基因调节的破坏会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。拟议的工作将为理解分子水平正常和患病状态的基因调节提供基础。这项工作的具体目的利用生化，结构和分子遗传学方法来检查两个激活域和两个共激活因子的直接靶标。我们将检查几种酸性激活剂互启动剂复合物的结构，以了解激活剂如何专门识别其靶标，激活剂 - 目标识别的常见原理以及更一般而言的本质上无序蛋白的功能。我们将检查传奇共激活因子与TBP（TATA结合蛋白）的相互作用，以及如何通过乙酰化调节这种相互作用。使用一组独特的交联试剂，我们将检查转录机械内介体共激活器复合物的直接靶标。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。结合在一起，我们的结果将导致一个分子模型，用于激活剂如何识别其共激活剂靶标，以及共激活因子如何通过与转录机械直接相互作用刺激基因表达。 公共卫生相关性：项目叙述这项研究的目的是了解转录的机制和调节，即mRNA合成过程。转录的调节是控制细胞生长，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。了解转录的机制及其调节将构成理解转录疾病中分子缺陷的基础，导致许多类型的癌症以及心脏病，神经系统疾病和先天缺陷。