Role of UTX condensation in chromatin regulation

UTX 缩合在染色质调控中的作用

• 批准号: 10365699
• 负责人: Hao Jiang
• 金额: $ 41.88万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10365699
• 关键词: Binding; Biochemical; Biological; Biological Assay; Biological Process; Biophysics; CRISPR/Cas technology; Cancer Biology; Cancer Cell Growth; Cell Differentiation process; Cells; ChIP-seq; Chimeric Proteins; Chromatin; Core Protein; Development; Developmental Process; Disease; EP300 gene; Engineering; Enhancers; Gene Expression; Genes; Genetic Transcription; Genomics; Histones; Homologous Gene; Human; Impairment; In Vitro; Kabuki Make-Up Syndrome; Liquid substance; Malignant Neoplasms; Mediating; Methylation; Methyltransferase; Missense Mutation; Molecular; Mus; Mutate; Mutation; Neurodevelopmental Disorder; Patients; Phase; Physical condensation; Physiological; Process; Property; Protein Dynamics; Proteins; Publications; RNA Splicing; Regulation; Role; Stem Cell Development; System; Transcription Initiation Site; Tumor Suppression; Tumor Suppressor Proteins; Variant; Work; Y Chromosome; biophysical analysis; biophysical properties; cancer cell; chromatin modification; demethylation; embryonic stem cell; genome editing; insight; mutant; neoplastic cell; novel; programs; reconstitution; recruit; stem cell differentiation; tumor; tumorigenesis

PROJECT ABSTRACT UTX/KDM6A is an important tumor suppressor and developmental regulator, and is frequently mutated in human cancers and certain neurodevelopmental disorders including Kabuki syndrome. Though UTX is a histone H3K27 demethylase, the demethylase activity is often dispensable in mediating tumor suppression and developmental regulation, and the key molecular activity of UTX in these processes remains elusive. UTX is thought to regulate chromatin activity by associating with and coordinating the function of MLL3/KMT2C and MLL4/KMT2D, the major H3K4 mono-methylase at enhancers, and the p300 histone acetyltransferase, all of which are also frequently mutated in human cancers. In our preliminary studies, we found that the tumor suppressive activity of UTX requires its phase separation property conferred by its core Intrinsically Disordered Region (cIDR). Endogenous UTX forms dynamic condensates in mouse embryonic stem cells (ESCs) and its cIDR is important for ESC differentiation. UTX and MLL4 form co-condensates that enhance the H3K4 mono-methylation activity. We also found that UTY, the Y chromosome homolog of UTX, has weaker tumor suppressive activity that is associated with formation of less dynamic condensates. These results let us formulate our central hypothesis that UTX needs to be in condensates with appropriate biophysical properties to be active in regulating gene expression on chromatin in regulating tumorigenesis and stem cell differentiation. We proposal two Specific Aims. Aim 1. Determine the mechanisms by which UTX condensation regulates chromatin activity. We will determine how UTX condensation regulates chromatin modification activities by biochemical reconstitution, and also through integrative analyses of multiple levels of chromatin activity in both transduced cancer cells and endogenous Utx-edited ESCs. Aim 2. Determine how biophysical properties of the UTX condensates regulate its biological activities. We will study biophysical properties of UTX WT and variant condensates and its associated co-activators. We will determine impacts of condensate properties on biological activities at different levels in in both transduced cancer cells and endogenous Utx-edited ESCs. We will also examine effect of disease-associated missense mutations of UTX on condensate properties.

项目摘要 UTX/KDM6A是重要的肿瘤抑制剂和发育调节剂，并且经常是 在人类癌症和包括Kabuki在内的某些神经发育障碍中突变 综合征。尽管UTX是组蛋白H3K27脱甲基酶，但脱甲基酶的活性通常是 可在介导肿瘤抑制和发育调控和关键分子方面可分配 在这些过程中UTX的活动仍然难以捉摸。认为UTX可以调节染色质活性 通过与MLL3/KMT2C和MLL4/KMT2D的功能相连并协调 H3K4单甲基酶在增强剂和p300组蛋白乙酰转移酶，所有这些都是 也经常在人类癌症中突变。在我们的初步研究中，我们发现肿瘤 UTX的抑制活动需要其核心赋予其相位分离属性 本质上无序区域（CIDR）。内源性UTX在小鼠中形成动态冷凝物 胚胎干细胞（ESC）及其CIDR对于ESC分化很重要。 UTX和MLL4 形式共同指示增强H3K4单甲基化活性。我们还发现了uty， Y UTX的Y染色体同源物具有较弱的肿瘤抑制活性，与之相关 形成较少的动态冷凝物。这些结果使我们提出了中心假设 该UTX需要在具有适当生物物理特性的冷凝物中活跃 调节染色质基因表达在调节肿瘤发生和干细胞分化中。 我们提出了两个具体目标。 AIM 1。确定UTX冷凝调节染色质的机制 活动。我们将通过 生化重构，以及通过多个染色质的综合分析 转导的癌细胞和内源性UTX编辑的ESC的活性。 目标2。确定UTX凝结物的生物物理特性如何调节其调节 生物活动。我们将研究UTX WT和变体冷凝物的生物物理特性 及其相关的共激活因子。我们将确定冷凝物特性对生物学的影响 转导的癌细胞和内源性UTX编辑的ESC的活性不同。 我们还将检查与疾病相关的UTX的错义突变对冷凝水的影响 特性。