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 基因组结构在基因活性控制中起着关键作用。在过去的几年里， 通过液-液相分离组装的无膜冷凝物已经成为一种 组织基因组的新主题，这是2018年科学突破之一。尽管 相分离对于基因组组织的重要性， 机制仍然是个谜。我们的总体目标是使用Polycomb组（PcG）蛋白作为 范例来理解液-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- 液相分离我们率先开发了体外和体内单分子 技术，使解决关键问题的基因组组织，通过液-液 相分离 基于这一概念和技术基础，我们的具体目标是确定 决定PcG缩合物在特定基因组位置成核的分子因素， 以确定H3 K27 me 3标记基因如何被携带的分子机制， 一起进入PcG缩合物，并确定PcG缩合物的功能作用， 反应动力学和转录抑制。我们将通过使用多- 学科方法，结合我们独特的单分子工具与生物化学，遗传， 和基因组分析。 我们的研究将建立一个范式转换模型，支持PcG蛋白如何 相分离以形成组织Polycomb靶基因以实现高效 生化反应和转录抑制。此外，我们将添加新的单分子 通过液-液相分离研究基因组组织的工具。因此，本研究将 对我们理解相分离如何组织 基因组，其影响远远超出表观遗传学领域。