Understanding mechanisms of transcriptional regulation by chromatin adaptor proteins

了解染色质接头蛋白的转录调控机制

• 批准号: 10533396
• 负责人: Yadira M Soto-Feliciano
• 金额: $ 24.9万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10533396
• 关键词: Adaptor Signaling Protein; Affect; Alleles; Autoimmunity; Award; Bar Codes; Binding; Biochemical; Biochemistry; Biological; Biological Assay; Biology; Cell physiology; Cells; ChIP-seq; Chemicals; Chromatin; Chromatin Remodeling Factor; Chromatin Structure; Clustered Regularly Interspaced Short Palindromic Repeats; Communities; Complex; DNA; DNA Binding; Data; Development; Disease; Distal; Endocrine; Enhancers; Environment; Epigenetic Process; Event; Functional disorder; Future; Gene Expression; Gene Expression Profiling; Genes; Genetic Transcription; Genome; Genomic approach; Genomics; Histones; Impairment; In Situ; Intellectual functioning disability; Intercistronic Region; Knowledge; Label; Language; Lead; Libraries; MLL gene; Malignant Neoplasms; Mediating; Menin; Mentors; Methods; Methyltransferase; Modification; Molecular; Motivation; Mutation; Nucleosomes; Oncogenic; Output; Phase; Phenotype; Physiological; Physiology; Play; Positioning Attribute; Post-Translational Protein Processing; Process; Proteins; Proteomics; Reader; Reading; Recurrence; Regulation; Regulatory Element; Research; Resources; Role; Scaffolding Protein; Shapes; Signal Transduction; Site; Surface; Testing; Tissues; Training; Transcriptional Regulation; Work; base; career; chromatin protein; chromatin remodeling; combinatorial; epigenomics; functional genomics; genetic approach; genome-wide; genomic locus; histone methyltransferase; histone modification; human disease; improved; innovation; insight; interest; leukemia; mutant; novel; pleiotropism; promoter; protein complex; protein structure; recruit; scaffold; structural biology; transcription factor; tumor

PROJECT SUMMARY Precise regulation of chromatin states is critical to many vital cellular processes, including differentiation and proliferation. Chromatin misregulation has been associated with human diseases, such as cancer, intellectual disabilities, and autoimmunity, among others. Understanding the mechanisms by which genomic access is controlled at the level of chromatin is critical for revealing how cellular phenotypes are established and maintained, and how these processes are altered in disease. This study focuses on elucidating the molecular mechanisms by which the chromatin adaptor Menin coordinates protein complexes to transduce chromatin signals into transcriptional outputs. This work will test the central hypothesis that context-specific functions of Menin are dictated by its ability to bind distinct chromatin environments and recruit a variety of chromatin factors in a highly regulated manner. During the mentored phase of the award, it will determine how Menin targets specific genomic loci by integrating DNA-barcoded nucleosome libraries, epigenomics, and functional genomics approaches. This multi-tiered approach will identify combinatorial histone modifications and transcription factors, as well as delineate their biological relationships with Menin and how they contribute to its genomic localization (Aim 1). It will also identify and comprehensively characterize chromatin proteins and complexes that mediate regulatory functions of Menin at distinct cis-regulatory elements (Aim 2). During the independent phase of the award, it will determine how recurrent MEN1 mutations affect Menin’s ability to target and shape the chromatin landscape, as well as affect transcription (Aim 3). Furthermore, it will characterize the cellular and organismal phenotypes elicited by endogenous expression of MEN1 mutant alleles. Successful completion of the proposed studies will produce insights into how Menin regulates chromatin biology and transcription, and will provide a greater understanding of the mechanisms by which disease-associated mutant Menin proteins promote disease. Such knowledge could yield novel insights into the roles of chromatin and epigenetic regulators in normal physiology and pathophysiology. The integrative approaches proposed here will serve as a valuable resource for the wider scientific community interested in pursuing future studies of chromatin factors with presumed adaptor/scaffolding functions that have not been studied previously due to limitations of current methods. They will also serve as a platform for the PI to obtain new training in chromatin and chemical biology, biochemistry of transcription, structural biology, and computational epigenomics. Such training will be critical for the development of the PI’s career and will position her to effectively integrate these approaches for making novel and innovative contributions to the field of chromatin biology.

项目概要 染色质状态的精确调节对于许多重要的细胞过程至关重要，包括分化和分化 增殖。染色质失调与人类疾病有关，例如癌症、智力障碍 残疾和自身免疫等。了解基因组访问的机制 在染色质水平上进行控制对于揭示细胞表型如何建立和 维持，以及这些过程如何在疾病中改变。本研究的重点是阐明分子 染色质接头 Menin 协调蛋白质复合物转导染色质的机制 信号转化为转录输出。这项工作将检验中心假设，即上下文特定的功能 Menin 取决于其结合不同染色质环境并招募各种染色质因子的能力 以高度监管的方式。在该奖项的指导阶段，它将确定梅宁的目标 通过整合 DNA 条形码核小体文库、表观基因组学和功能基因组学来确定特定基因组位点 接近。这种多层方法将识别组合组蛋白修饰和转录因子， 以及描述它们与 Menin 的生物学关系以及它们如何为其基因组定位做出贡献 （目标 1）。它还将识别并全面表征介导的染色质蛋白和复合物 Menin 在不同顺式调控元件上的调控功能（目标 2）。在独立阶段 该奖项将确定复发性 MEN1 突变如何影响 Menin 靶向和塑造染色质的能力 景观，以及影响转录（目标 3）。此外，它将表征细胞和有机体 MEN1 突变等位基因的内源表达引起的表型。顺利完成拟议的 研究将深入了解 Menin 如何调节染色质生物学和转录，并将提供 更好地了解与疾病相关的突变 Menin 蛋白促进疾病的机制。 这些知识可以对染色质和表观遗传调节因子在正常情况下的作用产生新的见解。 生理学和病理生理学。这里提出的综合方法将成为宝贵的资源 对于对未来染色质因子研究感兴趣的更广泛的科学界来说， 由于当前方法的限制，以前尚未研究过适配器/支架功能。他们 还将作为 PI 获得染色质和化学生物学、生物化学方面新培训的平台 转录、结构生物学和计算表观基因组学。此类培训对于发展至关重要 PI 职业生涯的一部分，并使她能够有效地整合这些方法来创造新颖和创新 对染色质生物学领域的贡献。