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.

项目摘要 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 增殖。染色质的正调与人类疾病（例如癌症）有关 残疾和自身免疫等。了解基因组访问的机制 在染色质水平上控制对于揭示细胞表型的建立和 维持以及这些过程如何改变疾病。这项研究的重点是阐明分子 染色质衔接子Menin将蛋白质复合物与染色质转导的机制 信号转录输出。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募集各种染色质因子的能力所决定的 以高度监管的方式。在奖励的修订阶段，它将决定梅宁的目标 特定的基因组基因座通过整合DNA-barcoded核小体库，表观基因组学和功能基因组学 方法。这种多层方法将确定组合的Hisstone修改和转录因子， 以及描述他们与梅宁的生物学关系以及它们如何为基因组定位做出贡献 （目标1）。它还将识别并全面地表征介导的染色质蛋白和复合物 Menin在不同顺式调节元件上的调节功能（AIM 2）。在独立阶段 奖励，它将决定复发性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（AIM 3）。此外，它将表征细胞和有机 Men1突变等位基因的内源性表达引起的表型。成功完成拟议的 研究将对Menin如何调节染色质生物学和转录产生见解，并将提供 对与疾病相关的突变蛋白促进疾病的机制有更深入的了解。 这种知识可以产生对染色质和表观遗传调节剂在正常作用的作用的新见解 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 对于有兴趣通过推测的染色质因素的研究感兴趣的更广泛的科学界 由于当前方法的局限性，适配器/脚手架函数先前尚未研究。他们 还将作为PI获得染色质和化学生物学的新培训的平台 转录，结构生物学和计算表观基因组学。这样的培训对于发展至关重要 PI的职业生涯，将使她有效地整合这些方法以制造新颖和创新的方法 对染色质生物学领域的贡献。