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.

项目摘要 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（例如组蛋白修饰剂，伴侣蛋白和染色质）介导 改建器。这些因素的突变与许多人类疾病密切相关。例如，突变 在保守的ATP依赖性染色质重塑的SWI/SNF家族中，与约20％的人有关 癌症。这些突变中的一些也与发展性和智力残疾综合症有关 作为棺材 - 西里斯综合征（CSS）。但是，我们不完全了解SWI/SNF重塑的哪些方面 活性在生理状况下的致病突变影响。重塑 染色质（RSC）复合物的结构是SWI/SNF家族的成员，也是唯一必需的 重塑酵母中的重塑。 RSC调节许多生物过程，包括所有三个生物学过程 RNA聚合酶。它至关重要地参与了在基因启动子附近维持规范的染色质结构。 在RSC ATPase亚基STH1中已经确定了许多域，这些域调节其重塑活性。 其他结构域与与DNA和核小体相互作用。但是， 这些领域指示活细胞中RSC功能的域知之甚少。此外，机制 调节RSC与染色质的关联也不清楚。 RSC可以与特定区域结合 染色质使用其在体外结合乙酰化组蛋白结合的溴结构域。 RSC如何 利用组蛋白乙酰化的招募或在生理条件下执行其功能 要理解。在特定目的中，使用酿酒酵母作为模型生物1），我们将 研究突变在各种调节和核小体结合结构域中的影响，以及一些 与染色质结构的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域中的突变如何影响细胞的能力 应对压力。在特定目标2）中，我们将确定促进RSC关联的组蛋白尾巴残基 与染色质和帮助RSC脱离染色质的染色质。乙酰化残基的程度 影响RSC使DNA访问的能力也将被确定，全基因组。此外，我们会的 检查RSC在调节伸长步骤中转录中的作用。这些研究在 了解组蛋白的修饰剂和染色质重塑如何合作调节基因表达。