Functional Analysis of Epigenetic Complexes

表观遗传复合物的功能分析

• 批准号: 10391329
• 负责人: ROBERT KINGSTON
• 金额: $ 87.68万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10391329
• 关键词: CHD7 gene; Cell Differentiation process; Cell Lineage; Cell Nucleus; Cells; Chromatin; Chromatin Structure; Complex; DNA; Development; Developmental Process; Epigenetic Process; Gene Expression Profile; Gene Expression Regulation; Genes; Human; Laboratories; Location; Mammalian Cell; Methylation; Mutation; Nucleosomes; Organ; Organism; PRC1 Protein; Polycomb; Proteins; Regulation; Regulator Genes; Repression; Role; Tissues; Work; appendage; base; cell behavior; cohesin; gene product; novel; protein function; zygote

Abstract: For humans to develop properly, the fertilized egg must divide in a manner that generates a body that has the correct organs and appendages in the correct places. Much of this is determined by the proper temporal and spatial regulation of master regulatory genes that guide formation of the differentiated cells that make up organs and appendages. A key aspect of this developmental process is the ability to maintain master regulatory genes in a repressed state in cells where their expression might cause inappropriate cell behavior. Mis-expression of even a single master regulatory gene, such as those encoded in the HOX loci, can result in a cell behaving in a manner incompatible with its body location and tissue type. The Polycomb- Group (PcG) group of genes is largely responsible for maintaining this repression, based upon intensive study over the past 70 years. Maintaining active expression patterns for genes is equally critical for development. A set of genes that maintains activation, called the trithorax- Group (trxG), was isolated via their ability to suppress PcG mutations. This application describes the continuation of our work on understanding the function of the proteins encoded by the PcG and trxG genes. These proteins form several complexes. The PcG complexes PRC1 and PRC2 are each large complexes that contain several PcG gene products. PRC1 is the main `engine' of repression in the Polycomb-Group and is known to interact with chromatin, the structure that packages genes in the nucleus of cells. Chromatin structure can render the enclosed DNA inaccessible to activating factors. A prominent hypothesis is that repression can be generated by generating highly packaged DNA that is no longer able to be transcribed and expressed. We describe approaches to investigate the various mechanisms used by PRC1 to generate packaged chromatin. PRC1 works together with PRC2 to generate repression. PRC2 methylates nucleosomes, the primary packaging unit of chromatin, on residue H3K27, a topic of intensive study in numerous laboratories and companies. We intend to provide novel information by exploring repressive activities of PRC2 that are unrelated to methylation. We will also characterize methylation by PRC2 of a separate protein involved in gene regulation, ElonginA. Finally, we will characterize two separate nucleosome remodeling activities encoded by trxG genes, the mammalian SWI/SNF (BAF) complex and mammalian CHD7. We will explore interactions between SWI/SNF subunits and cohesin, a chromatin organizing activity, and determine how those interactions impact long range interactions in chromatin. We will examine the role for a lncRNA called HERVH in targeting CHD7 activity.

摘要：要使人类正确发展，受精卵必须以一种方式分裂 生成一个在正确位置中具有正确器官和附属物的身体。其中很多 由主调节基因的适当时间和空间调节确定 指导组成器官和附属的分化细胞的形成。一个关键方面 这种发展过程的能力是在被压抑中维持主监管基因的能力 在其表达可能引起不适当细胞行为的细胞中的状态。错误表达 即使是单一的主调节基因，例如在HOX基因座中编码的基因，也可能导致细胞 表现方式与其身体位置和组织类型不兼容。 polycomb- 基于基因组（PCG）基因基在很大程度上负责维持这种抑制作用。 在过去的70年中，密集研究。维持基因的主动表达模式是 同样对发展至关重要。一组维持激活的基因，称为trithorax- 通过抑制PCG突变的能力来隔离组（TRXG）。此应用程序 描述了我们在理解编码蛋白质功能方面的工作的延续 由PCG和TRXG基因。这些蛋白质形成几种复合物。 PCG综合体PRC1 PRC2都是包含多种PCG基因产物的大型复合物。 PRC1是主要的 polycomb组中抑制的“发动机”，已知与染色质相互作用， 结构将基因包装在细胞核中。染色质结构可以使 封闭的DNA无法访问激活因子。一个明显的假设是镇压可以 通过生成不再能够转录的高度包装的DNA产生 表达。我们描述了研究PRC1使用的各种机制的方法 生成包装的染色质。 PRC1与PRC2一起生成抑制作用。 PRC2 残基H3K27的甲基元素核小体，染色质的主要包装单元，一个主题 在众多实验室和公司中进行深入研究。我们打算提供小说 通过探索与甲基化无关的PRC2抑制作用的信息。我们 还将通过PRC2表征涉及基因调节的单独蛋白的甲基化， Elongina。最后，我们将表征两个编码的两个单独的核小体重塑活性 由TRXG基因，哺乳动物SWI/SNF（BAF）复合物和哺乳动物CHD7。我们将 探索SWI/SNF亚基和粘蛋白之间的相互作用，染色质组织活性， 并确定这些相互作用如何影响染色质中的远距离相互作用。我们将 检查称为HERVH的LNCRNA在靶向CHD7活性中的作用。