Understanding mechanisms of transcriptional regulation by chromatin adaptor proteins

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是至关重要的，包括分化和分化。 增殖染色质失调与人类疾病有关，如癌症、智力障碍、 残疾和自身免疫等等。了解基因组获取的机制 在染色质水平上的控制对于揭示细胞表型是如何建立的至关重要， 以及这些过程在疾病中如何改变。这项研究的重点是阐明分子 染色质衔接子Menin协调蛋白质复合物与染色质结合的机制 信号转化为转录产物。这项工作将测试的中心假设，特定的功能， Menin是由它结合不同染色质环境和募集各种染色质因子的能力决定的 以高度规范的方式。在该奖项的指导阶段，它将决定如何Menin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， 接近。这种多层次的方法将识别组合的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有助于其基因组定位 (Aim 1）。它还将识别和全面表征染色质蛋白质和复合物，介导 Menin在不同的顺式调节元件上的调节功能（Aim 2）。在独立阶段， 该研究将确定MEN 1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 MEN 1突变等位基因的内源性表达引发的表型。圆满完成拟议的 研究将深入了解Menin如何调节染色质生物学和转录，并将提供一个新的研究方向。 更深入地了解疾病相关突变Menin蛋白促进疾病的机制。 这些知识可以产生新的见解染色质和表观遗传调节剂在正常细胞中的作用。 生理学和病理生理学。这里提出的综合办法将作为一种宝贵的资源 对于更广泛的科学界感兴趣的追求未来的研究染色质因子与假定 由于目前方法的限制，以前没有研究过适配器/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， 转录、结构生物学和计算表观基因组学。这种培训对于发展 PI的职业生涯，并将使她有效地整合这些方法，使新颖和创新 对染色质生物学领域的贡献。

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.