Structure and Mechanism of the SET1/COMPASS H3K4 Methyltransferase Complex

SET1/COMPASS H3K4 甲基转移酶复合物的结构和机制

• 批准号: 10667557
• 负责人: Champak Chatterjee
• 金额: $ 37.89万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-08 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10667557
• 关键词: ASH2L gene; Active Sites; Address; Animals; Binding; Biochemical; Biological; Biological Assay; Biological Models; Biology; Cells; Chemicals; Chromatin Structure; Complex; Congenital Heart Defects; Cryoelectron Microscopy; Data; Defect; Disease; Elements; Enhancers; Enzyme Kinetics; Enzymes; Epigenetic Process; Etiology; Eukaryota; Eukaryotic Cell; Family; Family member; Gene Expression; Gene Expression Regulation; Genetic; Genetic Transcription; HRX protein; Health; Histone H3; Histone-Lysine N-Methyltransferase; Histones; Human; In Vitro; Intellectual functioning disability; Light; Link; Lysine; MLL gene; MLL2 gene; Malignant Neoplasms; Mammals; Maps; Measures; Mental disorders; Methods; Methylation; Methyltransferase; Mixed-Lineage Leukemia; Molecular; Monoubiquitination; Motion; Mutation; Names; Nucleosomes; Orthologous Gene; Outcome; Post-Translational Protein Processing; Protein Subunits; Recombinants; Regulation; Reporting; Residual state; Role; Saccharomyces cerevisiae; Schizophrenia; Side; Signal Transduction; Specificity; Structure; Structure-Activity Relationship; Testing; Transcriptional Activation; Transcriptional Regulation; Work; Yeast Model System; Yeasts; autism spectrum disorder; clinically relevant; congenital heart disorder; enzyme substrate; epigenetic regulation; histone methylation; human disease; in vivo; interest; leukemia; member; methyl group; nervous system disorder; neuropsychiatric disorder; novel; paralogous gene; protein complex; prototype; reconstitution; response

Project Summary The post-translational modification of histone H3 lysine 4 (H3K4) by methyl groups is an evolutionarily conserved epigenetic mark that is generally associated with transcription activation in all eukaryotic cells. Early studies of the yeast model system, S. cerevisiae, have not only identified the prototype of the SET1/MLL family of methyltransferases as the enzyme responsible for H3K4 mono-, di-, and trimethylation, but also revealed a yeast Set1-centric protein complex, known as COMPASS, that stabilizes and confers catalytic activity to the enzyme. The SET1/MLL family of H3K4 methyltransferases has undergone a significant expansion in animals. Mammals have evolved a total of six distinct and functionally non-redundant family members, each of which also functions within a COMPASS or COMPASS-like complex. Remarkably, recent studies have shown that mutations or dysregulation of the six human SET1/MLL methyltransferases are associated with a spectrum of mental illnesses, including schizophrenia, autism, and intellectual disability disorders. Malfunctions of some of these family members are further linked to other human diseases such as mixed lineage leukemia and congenital heart disease. Despite their important biological roles and their high relevance to human health, a molecular and mechanistic understanding of the SET1/MLL H3K4 methyltransferases is largely lacking due to the large sizes of most SET1/MLL enzymes and the complexity associated with their assemblies and regulation. To date, most structural and biochemical studies have been focused on single domains and small fragments of the yeast and human SET1/MLL enzymes and COMPASS subunits. Many questions, such as how the SET1/MLL enzymes bind and become regulated by four common catalytic module subunits, namely RBBP5/Swd1, WDR5/Swd3, ASH2L/Bre2, and DPY30/Sdc1 (human/yeast ortholog), how the resulting complexes recognize H3K4 in the context of nucleosome and differentially catalyze mono- vs. multi-H3K4 methylation, and how the activities of COMPASS and COMPASS-like complexes are regulated by upstream signals such as H2B mono-ubiquitination remain unclear. Using a combination of structural, chemical and biochemical approaches, as well as yeast cell- based functional assays, we propose to dissect the structure and function relationship of the yeast Set1 COMPASS complex as a model system and extend this work to the clinically relevant human SET1/MLL complexes. Our proposed studies hold the promise to establish the missing framework for understanding the structural basis of the SET1/MLL H3K4 methyltransferase function and regulation in eukaryotic biology and unmasking the molecular mechanisms of various human diseases associated with their malfunction.

项目摘要 甲基的组蛋白H3赖氨酸4（H3K4）的翻译后修饰是一种进化保守的 通常与所有真核细胞中转录激活有关的表观遗传标记。早期研究 酵母模型系统S. cerevisiae不仅确定了set1/mll家族的原型 甲基转移酶是负责H3K4单，二甲基和三甲基化的酶，但也揭示了酵母 以SET1为中心的蛋白质复合物，称为指南针，可稳定并赋予酶的催化活性。 H3K4甲基转移酶的SET1/MLL家族在动物中发生了显着扩展。哺乳动物 总共进化了六个不同且功能性非冗余的家庭成员，每个家庭成员也起作用 在指南针或指南针般的复合体中。值得注意的是，最近的研究表明，突变或 六个人Set1/mll甲基转移酶的失调与精神范围有关 疾病，包括精神分裂症，自闭症和智力疾病。其中一些故障 家庭成员进一步与其他人类疾病有关，例如混合谱系白血病和先天性心脏 疾病。尽管它们具有重要的生物学作用及其与人类健康的高度相关性，但分子和 对SET1/MLL H3K4甲基转移酶的机械理解在很大程度上缺乏大小 大多数SET1/MLL酶及其组装和调节的复杂性。迄今为止，大多数 结构和生化研究集中在酵母的单个领域和小碎片上 人类set1/mll酶和指南针亚基。许多问题，例如SET1/MLL酶如何 绑定并受到四个常见催化模块亚基的调节，即rbbp5/swd1，wdr5/swd3， ASH2L/BRE2和DPY30/SDC1（人/酵母直系同源物），所产生的配合物如何识别H3K4 核小体和差异化催​​化单与多H3K4甲基化的背景，以及如何 指南针和指南针般的复合物受到上游信号（例如H2B单素化）的调节 保持不清楚。结构，化学和生化方法以及酵母细胞的结合 基于功能分析，我们建议剖析酵母set1的结构和功能关系1 指南针复合物作为模型系统，并将这项工作扩展到临床相关的人类set1/mll 复合物。我们拟议的研究有望建立缺失的框架来理解 SET1/MLL H3K4甲基转移酶功能和真核生物学和调节的结构基础 揭示与故障相关的各种人类疾病的分子机制。