Roles for chromatin remodeler RSC and histone acetyltransferases in regulating chromatin structure and transcription

染色质重塑剂 RSC 和组蛋白乙酰转移酶在调节染色质结构和转录中的作用

• 批准号: 10579529
• 负责人: Chhabi K Govind
• 金额: $ 42.98万
• 依托单位: OAKLAND UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-09 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10579529
• 关键词: ATP phosphohydrolase; Acetylation; Affect; Affinity; Animal Model; Auxins; Binding; Biological Process; Bromodomain; Cells; ChIP-seq; Chromatin; Chromatin Structure; Code; Coffin-Siris Syndrome; Complex; Coupled; DNA; DNA Polymerase II; DNA-Directed RNA Polymerase; Data; Development; Developmental Disabilities; Disease; Dissociation; Family; Family member; Fingers; Gene Expression; Genes; Genetic Transcription; Goals; Growth; HTATIP gene; Histone Acetylation; Histone H3; Histone H4; Histones; Human; In Vitro; Intellectual functioning disability; Length; Link; Lysine; Malignant Neoplasms; Mediating; Methods; Molecular; Molecular Chaperones; Mutation; N-terminal; Nucleosome Binding Domain; Nucleosomes; Open Reading Frames; Physiological; Positioning Attribute; Role; Running; Saccharomyces cerevisiae; Saccharomycetales; Sfh1; Slide; Stress; Surface Plasmon Resonance; Syndrome; TATA-Box Binding Protein; Tail; Transcription Elongation; Transcription Initiation Site; Uracil; Yeasts; chromatin remodeling; disease-causing mutation; genome-wide; histone acetyltransferase; histone modification; human disease; in vitro activity; in vivo; member; mutant; nuclease; prevent; promoter; public health relevance; recruit; transcriptome sequencing

PROJECT SUMMARY Dynamic changes to chromatin structure are essential for regulating gene expression in cells. These changes are mediated by chromatin-associated factors such as histone modifiers, chaperones, and chromatin remodelers. Mutations in these factors are strongly linked to many human diseases. For example, mutations in the conserved SWI/SNF family of ATP-dependent chromatin remodelers are linked to ~20% of human cancers. Some of these mutations are also linked to developmental and intellectual disability syndrome, such as Coffin-Siris syndrome (CSS). However, we do not fully understand what aspects of SWI/SNF remodeling activities are affected by the disease-causing mutations under physiological conditions. The Remodels the Structure of Chromatin (RSC) complex is a member of the SWI/SNF family, and is the only essential remodeler in budding yeast. RSC regulates many biological processes, including transcription by all three RNA polymerases. It is critically involved in maintaining canonical chromatin structure near gene-promoters. Many domains have been identified within the RSC ATPase subunit Sth1 that modulate its remodeling activity. Additional domains are implicated in interacting with DNA and nucleosomes. However, the contributions of these domains in dictating RSC function in living cells are poorly understood. Furthermore, the mechanisms that regulate the association of RSC with chromatin are also not clear. RSC could bind to specific regions of chromatin using its bromodomains that have been shown to bind acetylated histones in vitro. How RSC exploits histone acetylation for its recruitment or to execute its function under physiological conditions remains to be understood. Using Saccharomyces cerevisiae as a model organism, in the specific AIM 1), we will investigate the impact of mutations in various regulatory and nucleosome-binding domains, and some of the mutations that are linked to developmental abnormalities on chromatin structure, including accessibility and gene expression. We will examine how mutations in these important domains affect the ability of cells to respond to stress. In the specific AIM 2), we will identify the histone tail residues that promote RSC association with chromatin and those that help RSC disengage from chromatin. The extent to which acetylated residues affect RSC ability to make DNA accessible will also be determined, genome-wide. Furthermore, we will examine the role of RSC in regulating transcription during elongation steps. These studies will be valuable in understanding how histone modifiers and chromatin remodelers cooperate to regulate gene expression.

项目概要 染色质结构的动态变化对于调节细胞中的基因表达至关重要。这些变化 由染色质相关因子介导，例如组蛋白修饰剂、分子伴侣和染色质 改造者。这些因素的突变与许多人类疾病密切相关。例如，突变 在 ATP 依赖性染色质重塑的保守 SWI/SNF 家族中，与约 20% 的人类 癌症。其中一些突变还与发育和智力障碍综合症有关，例如 科芬-西里斯综合症（CSS）。然而，我们并不完全了解SWI/SNF重塑的哪些方面 生理条件下的致病突变会影响其活性。改造 染色质结构 (RSC) 复合体是 SWI/SNF 家族的成员，是唯一必需的 芽殖酵母中的重塑剂。 RSC 调节许多生物过程，包括所有三个过程的转录 RNA聚合酶。它对于维持基因启动子附近的规范染色质结构至关重要。 在 RSC ATP 酶亚基 Sth1 内已鉴定出许多调节其重塑活性的结构域。 其他结构域涉及与 DNA 和核小体的相互作用。然而， 人们对这些决定活细胞 RSC 功能的领域知之甚少。此外，机制 调节 RSC 与染色质关联的因素也尚不清楚。 RSC可以结合到特定区域 染色质使用其溴结构域，该结构域已被证明可以在体外结合乙酰化组蛋白。如何RSC 利用组蛋白乙酰化来招募或在生理条件下执行其功能 被理解。使用酿酒酵母作为模式生物，在具体的 AIM 1) 中，我们将 研究各种调控域和核小体结合域中突变的影响，以及一些 与染色质结构发育异常相关的突变，包括可及性和 基因表达。我们将研究这些重要领域的突变如何影响细胞的能力 对压力做出反应。在特定的AIM 2)中，我们将鉴定促进RSC关联的组蛋白尾部残基 与染色质以及帮助 RSC 脱离染色质的物质。乙酰化残基的程度 RSC 获取 DNA 的能力的影响也将在全基因组范围内确定。此外，我们将 检查 RSC 在延伸步骤中调节转录的作用。这些研究将具有价值 了解组蛋白修饰剂和染色质重塑剂如何合作调节基因表达。