Understanding mechanisms of transcriptional regulation by chromatin adaptor proteins

项目总结 染色质状态的精确调节对许多重要的细胞过程至关重要，包括分化和 扩散。染色质失调与人类疾病有关，如癌症、智力 残障和自身免疫等。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是如何建立的和 以及这些过程在疾病中是如何改变的。这项研究的重点是阐明分子 染色质适配器Menin协调蛋白质复合体转导染色质的机制 将信号转换成转录输出。这项工作将检验这一中心假设，即 以一种高度规范的方式。在奖项的指导阶段，它将决定梅宁如何瞄准 整合DNA条形码核小体文库、表观基因组学和功能基因组学的特定基因组位点 接近了。这种多层次的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对其基因组定位做出贡献 (目标1)。它还将识别和全面表征介导 Menin在不同顺式调节元件上的调节功能(目标2)。在独立的阶段， 获奖后，它将确定反复发生的MEN1突变如何影响梅宁靶向和塑造染色质的能力 景观，以及影响转录(目标3)。此外，它将描述细胞和生物体的特征 MEN1突变等位基因内源性表达引起的表型。圆满完成拟议中的 研究将深入了解梅宁如何调节染色质生物学和转录，并将提供一种 更好地理解与疾病相关的突变的Menin蛋白促进疾病的机制。 这些知识可以产生对染色质和表观遗传调节因子在正常 生理学和病理生理学。这里提出的综合方法将是一个宝贵的资源。 对于有兴趣进行染色质因子未来研究的更广泛的科学界来说，假设 由于当前方法的限制，以前没有研究过的适配器/脚手架功能。他们 还将作为一个平台，让PI获得关于染色质和化学生物学、生物化学的新培训 转录、结构生物学和计算表观基因组学。这样的培训将对发展至关重要 并将使她有效地整合这些方法，使其具有新颖性和创新性 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.