Role of Multimerization in H3.3/H4 deposition by the HIRA complex

多聚化在 HIRA 复合物沉积 H3.3/H4 中的作用

• 批准号: 10552580
• 负责人: Mary Szurgot
• 金额: $ 3.5万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-03 至 2024-03-02
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10552580
• 关键词: ASF1A gene; Aging; Area; Binding; Biochemical; Biological Assay; C-terminal; Calorimetry; Cell Differentiation process; Cell Nucleus; Cells; ChIP-seq; Chaetomium; Chromatin; Complement; Complex; Crowding; Cryoelectron Microscopy; DNA; DNA Binding; DNA Damage; Data; Deposition; Development; Dimerization; Disease; Environment; Epigenetic Process; Gene Expression; Genetic Materials; Genetic Transcription; Genome; Goals; Health; Hela Cells; Higher Order Chromatin Structure; Histone H3.3; Histones; Homodimerization; Human; In Vitro; Investigation; Knock-out; Label; Maintenance; Mediating; Modeling; Molecular; Molecular Chaperones; Mutate; Nuclear; Nucleosomes; Physiologic pulse; Play; Process; Protein Subunits; Proteins; Resolution; Rest; Role; Scaffolding Protein; Site; Site-Directed Mutagenesis; Structure; System; Titrations; UV induced DNA damage; Variant; Visualization; cell type; dimer; experimental study; fungus; hereditary prostate cancer protein 2; insight; meter; mutant; particle; protein complex; protein protein interaction; response; scaffold; thermophilic organism

Project Summary The goal of this project is to characterize, with molecular detail, the role that multimerization of the HIRA histone chaperone complex plays in depositing histone H3.3/H4 onto DNA. In order to fit roughly two meters worth of DNA into a ten-micron cell nucleus, histones are used as scaffolding proteins to condense linear genetic material into the higher order structures of chromatin. Epigenetic regulatory machinery is essential to ensure that the chromatin landscape is well maintained to allow for certain gene expressions to be readily activated or inactivated to regulate both cell fate and cell stability. The HIRA histone chaperone complex is responsible for delivering histone H3.3/H4 onto nascent DNA in areas of active gene transcription and is therefore implicated in chromatin changes during development, in maintenance of terminally differentiated cell types, and in aging. The strategies I propose will first provide understanding of the molecular basis of the interaction between the C-terminal trimerization domain of HIRA and CABIN1 and then will probe the relevance of these interactions in vitro by investigating the effect that mutants have on the histone deposition process using a nucleosome assembly assay and in cells by investigating the effect that mutants have on complex localization and histone deposition within the crowded nuclear landscape. The proposed study will deepen our understanding of a ubiquitous epigenetic mechanism essential for regulating the chromatin environment of healthy human cells.

项目摘要 这个项目的目标是用分子细节来表征多聚体的作用 Hira组蛋白伴侣蛋白复合体在将组蛋白H3.3/H4沉积到DNA上起作用。为了适应 大约相当于两米的DNA进入一个10微米的细胞核，组蛋白被用作支架 蛋白质将线性遗传物质浓缩成染色质的高阶结构。表观遗传 监管机制是确保染色质景观得到良好维护的关键，以允许 某些基因的表达很容易被激活或失活，从而调节细胞的命运和细胞 稳定性。Hira组蛋白伴侣复合体负责将组蛋白H3.3/H4传递到 活跃基因转录区域中的新生DNA，因此与染色质变化有关 在发育过程中，在终末分化细胞类型的维持中，在衰老中。这个 我提出的策略将首先提供对相互作用的分子基础的理解 Hira和CABIN1的C-末端三聚结构域，然后将探索它们的相关性 通过研究突变体对组蛋白沉积过程的影响进行体外相互作用 使用核小体组装试验并通过研究突变体对细胞的影响 拥挤的核景观中复杂的定位和组蛋白沉积。建议数 研究将加深我们对普遍存在的、对调控至关重要的表观遗传机制的理解 健康人体细胞的染色质环境。