Role of BAP1/ASXL3 complex in transcriptional regulation and development

BAP1/ASXL3 复合物在转录调控和发育中的作用

• 批准号: 10669750
• 负责人: Lu Wang
• 金额: $ 40万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-22 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10669750
• 关键词: Attenuated; Biochemical; Biological Markers; Bromodomain; Cells; Chromatin; Chromatin Structure; Complex; Development; Developmental Process; Disease; Enhancers; Epigenetic Process; Equilibrium; Functional disorder; Gene Expression; Genes; Genetic; Genetic Transcription; Goals; Histone H2A; Histones; Lysine; Malignant Neoplasms; Mammalian Cell; Mammals; Mediating; Modification; Molecular; Multiprotein Complexes; Mus; Mutation; Neurons; PRC1 Protein; Patients; Play; Polycomb; Prognostic Marker; Proteins; Role; Scaffolding Protein; Therapeutic; Transcription Coactivator; Transcriptional Regulation; Ubiquitination; developmental disease; driver mutation; embryonic stem cell; gene repression; histone modification; human disease; nerve stem cell; new therapeutic target; novel; novel therapeutics; recruit; screening; small molecule; small molecule inhibitor; stem cell differentiation; transcriptome; ubiquitin isopeptidase

Project Summary Recent advances have shown that dysfunctions prevalent in epigenetic factors play an important role in the developing human diseases, including developmental disorders and cancers. Therefore, understanding the roles of these epigenetic factors may aid in identifying new epigenetic prognostic markers or targetable biomarkers that could contribute towards the establishment of novel therapeutics. Histone H2A lysine 119 ubiquitination (H2AK119ub) is one of the most functional histone marks that plays an essential role in establishing repressive chromatin domains and mediating Polycomb induced transcriptional repression during development. The evolutionarily conserved H2AK119ub histone modification is mainly catalyzed by the Polycomb repressive complex 1 (PRC1) and deubiquitinated by Polycomb repressive-deubiquitinase (PR-DUB) complex (also known as the BAP1 complex in mammals) that antagonizes PRC1’s function. In mammalian cells, the BAP1 complex functions as a multi-protein complex, containing as many as ten different subunits, which are responsible for the chromatin recruitment, protein stability, and enzymatic activity of BAP1. Mutations and dysregulations within subunits in the BAP1 complex are found in patients with developmental diseases, neuronal disorders, and cancer. Therefore, it is critical to understand the molecular basis of how genes are turned on/off by the BAP1/PRC1 epigenetic balance. Our previous studies have characterized a functional epigenetic axis comprised of both BAP1 and Bromodomain and Extraterminal (BET)-containing protein 4 (BRD4), which are physically bridged together by the scaffold protein ASXL3—the largest subunit within the BAP1 complex. Genetic depletion of the linker, ASXL3, dramatically attenuates the establishment of the BAP1/ASXL3/BRD4 epigenetic axis machinery at active enhancers, leading towards a significant reduction in the enhancer-nearby gene expression. In our current studies, we have employed biochemical, molecular, and small-molecule screening approaches to mechanistically understand how the ASXL3/BAP1/BRD4 epigenetic axis regulates transcription and determines cell fate and differentiation ability. Our first goal for this study is to uncover the BAP1/ASXL3 sub-complex’s role at active enhancers and assess the impact of the BAP1/ASXL3/BRD4 epigenetic axis on enhancer activity, chromatin structure, and gene expression. As a major H2AK119 deubiquitinase, the BAP1 complex functions as a general transcriptional activator, antagonizing PRC1’s function and is involved in PRC1-dependent transcriptional regulation. Therefore, our second project is to elucidate the relationship between BAP1 and a distinct PRC1 sub-complex in regulating H2AK119ub levels by utilizing our newly developed BAP1-specific small-molecule inhibitor. Mutations within ASXL3 have been demonstrated to be driver mutations in multiple neuronal diseases. Therefore, we will define the role of ASXL3 in mediating ESC differentiation into neuronal progenitor cells (NPC), and then investigate how ASXL3 and its associated epigenetic factors determine the transcriptome landscape during developmental processes.

项目摘要 最近的进步表明，表观遗传因素中普遍存在的功能障碍在 发展人类疾病，包括发育障碍和癌症。因此，了解角色 在这些表观遗传因素中，可能有助于识别新的表观遗传预后标记或可靶向的生物标志物 这可能有助于建立新的疗法。组蛋白H2A赖氨酸119泛素化 （H2AK119UB）是功能最强的组蛋白标记之一，在建立反射性方面起着至关重要的作用 染色质结构域和介导PolyComb在发育过程中诱导转录表达。这 进化配置的H2AK119UB HISSTONE修饰主要由Polycomb反射性催化 复合物1（PRC1）和由Polycomb反射泛素酶（PR-DUB）复合物（也已知）去泛素化 作为哺乳动物中的BAP1复合物）与PRC1功能的拮抗。在哺乳动物细胞中，BAP1复合物 充当多蛋白络合物，包含多达十个不同的亚基，这是负责的 BAP1的染色质募集，蛋白质稳定性和酶活性。内部的突变和失调 BAP1复合物中的亚基在发育疾病，神经元疾病和 癌症。因此，了解基因如何打开/关闭的分子基础是至关重要的 BAP1/PRC1表观遗传平衡。我们以前的研究表征了一个功能性表观遗传轴完成 BAP1和溴结构域和含有含蛋白4（BRD4）的溴化胺和外部溴化胺（BET），它们在物理上是 由脚手架蛋白ASXL3（BAP1复合物中最大的亚基）桥接在一起。遗传部署 链接器的ASXL3极大地削弱了BAP1/ASXL3/BRD4表观遗传轴的建立 活跃增强剂的机械，导致增强子 - 核基因表达的显着降低。 在目前的研究中，我们采用了生化，分子和小分子筛选方法 机械学上了解ASXL3/BAP1/BRD4表观遗传轴如何调节转录并确定 细胞命运和分化能力。这项研究的第一个目标是揭示BAP1/ASXL3子复合物的角色 在主动增强器中，评估BAP1/ASXL3/BRD4表观遗传轴对增强剂活性的影响， 染色质结构和基因表达。作为主要的H2AK119去泛素酶，BAP1复合物充当 一般的转录激活剂，拮抗Prc1的功能，并参与PRC1依赖性 转录调节。因此，我们的第二个项目是阐明BAP1和A之间的关系 通过使用我们新开发的BAP1特异性 小分子抑制剂。已证明ASXL3内的突变是多个驱动器突变 神经元疾病。因此，我们将定义ASXL3在将ESC分化为神经元中的作用 祖细胞（NPC），然后研究ASXL3及其相关的表观遗传因素如何确定 发育过程中的转录组景观。