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

基本信息

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
项目类别：
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

项目摘要

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.

项目概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、认知和有意识的思维。这些大量的细胞都起源于相对较小的神经祖细胞池，这些神经祖细胞进行分裂和分裂胚胎发育过程中根据时空线索进行分化。正确的发展这些细胞的功能需要极其精确的基因表达控制，这是由两者介导的转录因子和表观遗传修饰因子。虽然转录因子的突变是好的其特征是导致神经发育缺陷，但对其机制知之甚少表观遗传修饰突变会导致神经发育缺陷。该项目将开始填补这一空白分析 Polycomb 抑制复合物 2 (PRC2) 复合物，这是一种甲基化的表观遗传修饰剂组蛋白 H3 (H3K27me3) 的赖氨酸 27，调节 RNA 聚合酶 II 转录，并且对于神经祖细胞的形成和功能。具体来说，我的项目将调查 PRC2 的监管神经祖细胞增殖和分化为神经元时的功能和基因靶向命运。使用免疫沉淀-质谱法，我们最近发现了一种新的相互作用 PRC2 成分和一个关键的神经发生决定因素，它可能具有从未被发现的功能基因调控。我们将使用多学科方法来确定这一机制的细节相互作用并阐明这些相互作用如何影响 PRC2 功能。为了实现这个目标，我们首先要生化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 PRC2 与基因靶标的结合并产生影响转录活动。最后，我们将描述这种相互作用对自我更新和神经祖细胞的分化能力。我们的研究结果将揭示分子机制关键的神经发生决定因子与 PRC2 相互作用，以编程早期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的机制调节早期神经发育。我研究的长期目标是定义染色质修饰剂和转录回路在发育环境中调节基因表达并确定这种串扰的功能障碍如何导致人类出生缺陷和疾病。