Mechanisms of eukaryotic transcription activation

真核转录激活机制

• 批准号: 8770495
• 负责人: Steven M Hahn
• 金额: $ 54.57万
• 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 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8770495
• 关键词: Address; Binding; Biochemical; Biochemistry; Complex; Computer Analysis; Congenital Abnormality; Defect; Development; Diagnosis; Disease; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Goals; Grant; Growth and Development function; Heart Diseases; Human; Individual; Length; Mammalian Cell; Mass Spectrum Analysis; Mediator of activation protein; Methods; Modeling; Molecular; Molecular Conformation; Molecular Genetics; Outcome; Outcome Study; Pathway interactions; Physiological; Property; Proteins; RNA Polymerase II; Reagent; Recruitment Activity; Regulation; Regulator Genes; Research; Saccharomyces cerevisiae; Sampling; Signal Pathway; Specificity; Sum; Syndrome; System; Technology; Testing; Therapeutic Intervention; Transcription Coactivator; Transcription Elongation; Transcription Initiation; Transcriptional Activation; Transcriptional Regulation; Work; Yeasts; base; biological adaptation to stress; biological systems; cancer type; cell growth; gene correction; genetic regulatory protein; high throughput screening; human disease; molecular recognition; nervous system disorder; novel; novel strategies; promoter; protein crosslink; protein function; protein protein interaction; public health relevance; therapy design; therapy development

DESCRIPTION (provided by applicant): Summary Transcription activation, a key step in gene control, is the readout of many signaling pathways controlling cell growth, development and stress response and defects in activation cause many human diseases and syndromes. Activation typically results from factors containing activation domains (ADs) binding to coactivator complexes containing activator binding domains (ABDs). Most known ADs are unstructured in the absence of a binding partner and many interact with multiple and structurally-unrelated ABDs. The mechanisms used by ADs to interact with coactivators, the specificity of these interactions, and mechanisms of coactivator cooperativity are critical unanswered questions for understanding the molecular basis of gene regulation. The long-term goal of this project is to determine mechanisms used by gene-specific activators and coactivators to regulate RNA polymerase (Pol) II transcription. The objectives of this proposal are to determine: mechanisms used by ADs to recognize their coactivator targets, cooperative mechanisms used by the coactivators Mediator and TFIID, and what constitutes a functional AD and ABD. This work will utilize an interdisciplinary combination of biochemical, structural, molecular, and computational approaches to examine activation in S. cerevisiae. These mechanisms are likely to be conserved, since nearly all activators tested function in both yeast and mammalian cells. To understand these mechanisms, we will build upon several breakthrough concepts and methods developed in the past grant period. We will use protein crosslinking and mass spectrometry, combined with state-of-the-art NMR to examine mechanisms used by broadly-acting ADs when binding their coactivator targets - in both large physiologically relevant complexes and at the atomic level. We will examine cooperativity between Mediator and TFIID, two conserved and widely used transcription coactivators. Finally, we will use a combination of molecular, structural, and computational approaches to delineate what comprises AD function. These new approaches will reveal mechanisms of activator binding and specificity, mechanisms of coactivator cooperation, and find rules that explain a large class of broadly acting ADs. Our proposed research is significant because it will reveal new molecular recognition mechanisms that are important for understanding transcriptional regulation as well as a wide variety of interactions involving inherently disordered proteins that function in many biological systems.

描述（由申请人提供）： 摘要 转录激活是基因控制的关键步骤，是控制细胞生长、发育和应激反应的许多信号通路的读出，激活缺陷会导致许多人类疾病和综合症。激活通常是由含有激活结构域 (AD) 的因子与含有激活剂结合结构域 (ABD) 的共激活剂复合物结合所致。大多数已知的 AD 在没有结合伴侣的情况下都是非结构化的，并且许多 AD 与多个结构不相关的 ABD 相互作用。 AD 与共激活因子相互作用的机制、这些相互作用的特异性以及共激活因子协同性的机制是理解基因调控的分子基础的关键的未解答的问题。该项目的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 (Pol) II 转录的机制。该提案的目标是确定：AD 用于识别其共激活因子目标的机制、共激活因子 Mediator 和 TFIID 使用的合作机制，以及功能性 AD 和 ABD 的构成。这项工作将利用生物化学、结构、分子和计算方法的跨学科组合来检查酿酒酵母的激活。这些机制可能是保守的，因为几乎所有激活剂都在酵母和哺乳动物细胞中测试了功能。为了理解这些机制，我们将基于过去资助期间开发的几个突破性概念和方法。我们将使用蛋白质交联和质谱分析，结合最先进的 NMR 来检查广泛作用的 AD 在结合其共激活剂靶标时所使用的机制 - 在大型生理相关复合物中和原子水平上。我们将研究 Mediator 和 TFIID 这两种保守且广泛使用的转录共激活因子之间的协同作用。最后，我们将结合分子、结构和计算方法来描述 AD 功能的组成部分。这些新方法将揭示激活剂结合和特异性的机制、共激活剂合作的机制，并找到解释一大类广泛作用的 AD 的规则。我们提出的研究意义重大，因为它将揭示新的分子识别机制，这对于理解转录调控以及涉及在许多生物系统中发挥作用的固有无序蛋白质的各种相互作用非常重要。