Spatial Organization of Epigenetic Complexes and Chromatin

表观遗传复合物和染色质的空间组织

• 批准号: 10254304
• 负责人: Xiaojun Ren
• 金额: $ 31.65万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-18 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10254304
• 关键词: 3-Dimensional; AT-Hook Motifs; Address; Affect; Binding; Biochemical Genetics; Biochemical Reaction; Biological Assay; Biological Models; Cell Line; Cell physiology; Cells; Cellular biology; Chromatin; Complement; Complex; Coupled; DNA Binding; DNA sequencing; Development; Developmental Gene; Diffusion; Drosophila genus; Elements; Epigenetic Process; Filament; Foundations; Genes; Genetic Transcription; Genome; Genomics; Goals; Head; Homologous Gene; In Vitro; Kinetics; Knockout Mice; Life; Liquid substance; Location; Malignant Neoplasms; Microscopy; Modeling; Molecular; Mus; Mutation; PRC1 Protein; Phase; Physiology; Play; Polycomb; Polyhomeotic; Proteins; Reaction; Repression; Research; Resolution; Role; Science; Seeds; Tail; Techniques; Time; Transcriptional Regulation; base; embryonic stem cell; gene repression; in vivo; innovation; insight; interdisciplinary approach; intermolecular interaction; mutant; next generation; polymerization; recruit; single molecule; tool; transcriptome sequencing

Project Summary The genome organization plays a critical role in control of gene activity. Over the past few years, membraneless condensates assembled through liquid-liquid phase separation have emerged as a new theme in organizing the genome, which is one of science's 2018 breakthroughs. Despite the importance of phase separation for the genome organization, the underlying molecular mechanisms remain enigmatic. Our general goal is to use Polycomb group (PcG) proteins as a paradigm to understand how liquid-liquid phase separation organizes the genome. We recently demonstrated that PcG proteins assemble into condensates through liquid- liquid phase separation. We have pioneered the development of in vitro and in vivo single-molecule techniques that enable addressing key questions in the genome organization through liquid-liquid phase separation. Based upon this conceptual and technical foundation, our specific goal is to identify the molecular factors that dictate the nucleation of the PcG condensates at specific genomic locations, to define the molecular mechanism that underpins how H3K27me3-marked genes are brought together into the PcG condensates, and to determine the functional roles of PcG condensates in reaction kinetics and transcriptional repression. We will achieve this goal by using a multi- disciplinary approach that combines our unique single-molecule tools with biochemical, genetic, and genomic analyses. Our studies will establish a paradigm-shifting model that underpins how PcG proteins phase separate to form condensates that organize Polycomb target genes to achieve efficient biochemical reactions and transcriptional repression. Moreover, we will add new single-molecule tools to study the genome organization through liquid-liquid phase separation. Thus, this study will have a deep and sustained impact on our understanding of how phase separation organizes the genome, which has implications far beyond the epigenetic field.

项目摘要 基因组组织在控制基因活性中起着关键作用。在过去的几年中， 通过液体液相分离组装的膜无凝结物已作为A出现 组织基因组的新主题，这是科学2018年的突破之一。尽管有 相位分离对基因组组织的重要性，即基础分子 机制仍然神秘。我们的一般目标是将PolyComb组（PCG）蛋白作为一个 范式了解液 - 液相分离如何组织基因组。 我们最近证明了PCG蛋白通过液体组装成冷凝物 液相分离。我们开创了体外和体内单分子的发展 通过液体液体来解决基因组组织中的关键问题的技术 相分离。 基于这个概念和技术基础，我们的具体目标是确定 决定PCG在特定基因组位置凝结成核的分子因子， 定义基于H3K27me3标记基因的分子机制 一起进入PCG凝结物，并确定PCG凝结物的功能作用 反应动力学和转录抑制。我们将通过使用多种多样来实现这一目标 纪律方法将我们独特的单分子工具与生化，遗传， 和基因组分析。 我们的研究将建立一个基于PCG蛋白的范式转移模型 相分开以形成组织多肉液靶基因以实现有效的冷凝水 生化反应和转录抑制。此外，我们将添加新的单分子 通过液体液相分离研究基因组组织的工具。因此，这项研究将 对我们对相位分离如何组织的理解有深远和持续的影响 基因组的含义远远超出了表观遗传场。