Molecular Mechanisms Underlying the Regulation of Local Chromatin Structure

局部染色质结构调控的分子机制

• 批准号: 10430063
• 负责人: Sheena D'Arcy
• 金额: $ 38.25万
• 依托单位: UNIVERSITY OF TEXAS DALLAS
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10430063
• 关键词: Antifungal Agents; Antineoplastic Agents; Binding Proteins; Biochemistry; Biological Assay; Biological Process; Cells; Chromatin; Chromatin Structure; Complex Analysis; DNA; DNA Repair; DNA Sequence; DNA biosynthesis; Deuterium; Development; Disease; EP300 gene; Gene Expression Profile; Genetic Transcription; Genomic DNA; Histone H1; Histones; Human; Hydrogen; Lead; Malignant Neoplasms; Molecular; Molecular Chaperones; Nucleosomes; Pathogenicity; Play; Post-Translational Protein Processing; Regulation; Research; Resolution; Role; System; Therapeutic; Variant; Work; Yeasts; anticancer treatment; high throughput screening; histone acetyltransferase; histone modification; in vivo; interest; novel; public health relevance; translational impact; translational study

PROPOSAL SUMMARY The packaging of genomic DNA into repeating nucleosome units plays a pivotal role in the regulation of essential biological process including DNA transcription, DNA replication, and DNA repair. Cells must assemble and disassemble nucleosomes to read the information stored in the DNA sequence in a temporally and spatially regulated way. As such, cells have evolved several mechanisms that precisely regulate nucleosome dynamics and influence the local structure of chromatin. These mechanisms include: (1) the incorporation of histone variants into the nucleosome, (2) post-translational modification of histones, (3) histone chaperone activity, (4) ATP-dependent nucleosome remodeling, and (5) nucleosome-binding proteins. We currently do not know how these mechanisms act synergistically, nor how they influence intrinsic nucleosome dynamics. The research proposed in this MIRA application will focus on the quantification of nucleosome dynamics and the molecular basis of its regulation. We will develop a high-throughput assay for the quantification of nucleosome dynamics that will be applied to nucleosome assembly intermediates, mono-nucleosomes and multi- nucleosome arrays. This assay revolves around hydrogen-deuterium exchange analysis chromatin substrates in varied biologically-relevant states. We will also characterize the molecular mechanisms underpinning concerted activity of histone chaperones and histone acetyltransferases. Systems of interest at Rtt109, Vps75 and Asf1; as well as Nap1, CBP/p300 and linker histone H1. Varied approaches will be employed including biochemistry, high-resolution structural characterization, and in vivo complex analysis. The proposed work will impact translational studies motivated to develop anti-fungal therapeutics and identify new avenues for targeting of anti-cancer treatments.

提案摘要 基因组DNA包装成重复的核小体单位在调节细胞凋亡中起着关键作用。 重要的生物过程包括DNA转录、DNA复制和DNA修复。细胞必须组装 并拆解核小体，以在时间和空间上读取存储在DNA序列中的信息。 规范的方式。因此，细胞已经进化出多种精确调节核小体动力学的机制 并影响染色质的局部结构。这些机制包括：（1）组蛋白的掺入 变体进入核小体，（2）组蛋白的翻译后修饰，（3）组蛋白伴侣活性，（4） ATP依赖的核小体重塑;（5）核小体结合蛋白。我们目前还不知道 这些机制协同作用，也不知道它们如何影响内在核小体动力学。 MIRA申请中提出的研究将集中在核小体动力学的量化上 及其调控的分子基础。我们将开发一种高通量检测方法，用于定量 核小体动力学，将适用于核小体组装中间体，单核小体和多核小体， 核小体阵列该分析围绕氢-氘交换分析染色质底物 在不同的生物学相关状态下。我们还将描述支持 组蛋白伴侣和组蛋白乙酰转移酶的协同活性。在Rtt 109，Vps 75上感兴趣的系统 和Asf 1;以及Nap 1、CBP/p300和连接体组蛋白H1。将采用各种方法，包括 生物化学、高分辨率结构表征和体内复合物分析。拟议的工作将 影响转化研究，旨在开发抗真菌疗法并确定靶向治疗的新途径 抗癌治疗的方法。