Elucidation of the PRC2 interactome in early human neural development

阐明早期人类神经发育中的 PRC2 相互作用组

• 批准号: 9397068
• 负责人: Myron K Evans
• 金额: $ 5.71万
• 依托单位: ST. JUDE CHILDREN'S RESEARCH HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-02-01 至 2021-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9397068
• 关键词: Acetylation; Affect; Alpha Cell; Amino Acids; Binding; Biochemical; Biological Assay; Brain; Cell Count; Cell Differentiation process; Cell Maintenance; Cell Nucleus; Cell membrane; Cell physiology; Chromatin; Chromatin Structure; Cognition; Complex; Congenital Abnormality; Conscious; Cytoplasm; DNA; DNA Polymerase II; DNA-Binding Proteins; Data; Defect; Detection; Development; Developmental Gene; Disease; Embryonic Development; Epigenetic Process; Esthesia; Etiology; Functional disorder; Gene Expression; Gene Expression Regulation; Gene Targeting; Genetic Transcription; Genome; Genomics; Goals; Histone H3; Histones; Human; Immunoprecipitation; In Vitro; Knowledge; Lead; Link; Lysine; Mass Spectrum Analysis; Mediating; Methylation; Methyltransferase; Mission; Modification; Molecular; Mutagenesis; Mutation; Neurons; Nuclear; Output; PRC1 Protein; Phosphorylation; Physiological; Play; Pluripotent Stem Cells; Polycomb; Prevention; Process; Proliferating; Property; Proteins; RNA; RNA Polymerase II; Recruitment Activity; Regulation; Research; Role; Scientist; Signal Pathway; Signaling Molecule; Site; Stimulus; Synapses; Technical Expertise; Tertiary Protein Structure; Testing; United States National Institutes of Health; Writing; burden of illness; cell type; daughter cell; histone modification; in vivo; insight; interdisciplinary approach; mind control; mutant; nerve stem cell; nervous system disorder; neurodevelopment; neurogenesis; neuron development; novel; numb protein; programs; response; self-renewal; spatiotemporal; stem cells; tool; trait; transcription factor

Project Summary My research investigates the mechanisms by which epigenetic modifiers regulate the developing mammalian brain. The mammalian brain contains well over 1010 neurons, which synapse together to perform multiple functions including but not limited to sensation, cognition, and conscious thought. These vast numbers of cells all originate from a relatively small pool of neural progenitors, which divide and differentiate in response to spatiotemporal clues during embryonic development. The correct development and function of these cells requires exquisitely accurate gene expression control that is mediated by both transcription factors and epigenetic modifiers. While mutations in transcription factors are well characterized for causing neurodevelopmental defects, little is known about the mechanisms by which epigenetic modifier mutations cause neurodevelopmental defects. This project will begin filling this gap by analyzing the Polycomb Repressive Complex 2 (PRC2) complex, an epigenetic modifier that methylates lysine 27 of histone H3 (H3K27me3), regulates RNA polymerase II transcription, and is necessary for neural progenitor formation and function. Specifically, my project will investigate the regulation of PRC2 function and gene targeting within neural progenitor cells as they proliferate and differentiate to neuronal fates. Using immunoprecipitation-mass spectrometry, we recently identified a novel interaction between a PRC2 component and a key neurogenesis determinant, which likely has a never-discovered function in gene regulation. We will use multidisciplinary approaches to establish the mechanistic details of this interaction and to elucidate how these interactions affect PRC2 function. To achieve this goal, we will first biochemically identify the protein domains or amino acids required for this molecular interaction. Next, we will determine how this neurogenesis determinant affects PRC2 binding to gene targets and influences transcription activities. Finally, we will delineate the effect of this interaction on the self-renewal and differentiation capacity of neural progenitors. Our findings will reveal the molecular mechanism by which a crucial neurogenesis determinant interacts with PRC2 to program gene expression required for early neurogenesis. My research uses multiple tools to elucidate mechanisms by which epigenetic modifiers regulate early neural development. The long-term goal of my studies is to define the cross-talk between chromatin modifiers and transcriptional circuitry in regulating gene expression in developmental contexts and to determine how dysfunction in this cross-talk contributes to human birth defects and diseases.

项目摘要 我的研究调查了表观遗传修饰剂调节发育的机制， 哺乳动物的大脑哺乳动物的大脑包含超过1010个神经元，这些神经元通过突触连接在一起， 执行多种功能，包括但不限于感觉、认知和有意识的思考。这些 大量的细胞都起源于一个相对较小的神经祖细胞库，这些神经祖细胞分裂并 在胚胎发育过程中对时空线索的反应是不同的。正确的发展 这些细胞的功能需要精确的基因表达控制， 转录因子和表观遗传修饰因子。转录因子的突变 其特征是导致神经发育缺陷，关于其机制知之甚少， 表观遗传修饰突变导致神经发育缺陷。该项目将开始填补这一空白， 分析多梳抑制复合物2（PRC 2）复合物，一种表观遗传修饰剂， 组蛋白H3的赖氨酸27（H3 K27 me 3），调节RNA聚合酶II的转录，并且是 神经祖细胞的形成和功能。具体来说，我的项目将研究PRC 2的监管 当神经祖细胞增殖和分化为神经元细胞时， 命运使用免疫沉淀-质谱，我们最近发现了一种新的相互作用， PRC 2成分和一个关键的神经发生决定因素，可能在神经发生中具有从未发现的功能。 基因调控我们将使用多学科的方法来建立这一机制的细节， 相互作用，并阐明这些相互作用如何影响PRC 2功能。为了实现这一目标，我们将首先 生物化学鉴定这种分子相互作用所需的蛋白质结构域或氨基酸。接下来我们 将确定这种神经发生决定因素如何影响PRC 2与基因靶点的结合， 转录活动。最后，我们将描述这种相互作用对自我更新的影响， 神经祖细胞的分化能力。我们的研究结果将揭示一种 关键的神经发生决定子与PRC 2相互作用，以编程早期神经发生所需的基因表达。 神经发生我的研究使用多种工具来阐明表观遗传修饰剂 调节早期神经发育。我研究的长期目标是确定 在发育环境中调节基因表达的染色质修饰物和转录回路 并确定这种串扰中的功能障碍如何导致人类出生缺陷和疾病。