Understanding the Role of Combinatorial Histone PTM Patterns

描述(申请人提供)：组蛋白的翻译后修饰(PTM)构成了真核生物中调节染色质结构和基因表达模式的一种主要表观遗传机制。除了它们在标准生理学中的重要作用外，组蛋白PTM信号模式的中断被认为是各种人类疾病(如癌症)的重要、潜在的致病因素。由于染色质生物学领域中的大多数组蛋白PTM工作都是使用位点特异性抗体来完成的，因此对同一分子上共同存在的组合组蛋白PTM的定量测量一直没有得到满足。我们的目标包括继续开发基于质谱学的蛋白质组学和生物信息学方法来定量询问组合组蛋白PTM模式，并应用这些方法来研究组蛋白介导的表观遗传学机制，其背后是与健康相关的生物学研究的关键领域。在这里，我们将特别应用我们的方法来研究人类胚胎干细胞分化过程中表观遗传学的组蛋白PTM信号。我们的具体目标有三个，包括识别干细胞分化过程中组蛋白PTM的变化，鉴定翻译这些PTM模式的组蛋白PTM结合蛋白复合体，以及确定这些组合PTM在维持多能性状态或促进特定谱系形成中的作用。我们预计，这些综合的蛋白质组策略将继续产生新的工具来研究表观遗传学的组蛋白PTM，并对组合组蛋白PTM在不同生物事件(如细胞分化)的基因调控中的机制产生新的见解。

DESCRIPTION (provided by applicant): Post-translational modifications (PTMs) to histone proteins constitute a major type of epigenetic mechanism that regulates chromatin structure and gene expression patterns in eukaryotes. In addition to their important roles in standard physiology, disruptions in histone PTM signaling patterns have been suggested to be significant, potentially causative factors in various human diseases such as cancer. As most histone PTM work in the chromatin biology field is accomplished using site-specific antibodies, the quantitative measurement of combinational histone PTMs co-occurring on the same molecule has been unmet. Our objectives include the continued development of mass spectrometry-based proteomics and bioinformatic methods for quantitatively interrogating combinatorial histone PTM patterns, and applying these approaches to investigate histone mediated epigenetics mechanisms behind key areas of health related biological research. Here we will specifically apply our approaches to investigate epigenetic histone PTM signaling during human embryonic stem cell differentiation. Our specific aims are three in number and include identifying changing histone PTMs during stem cell differentiation, characterizing combinatorial histone PTM binding protein complexes that translate these PTM patterns, and determining the role of these combinatorial PTMs in maintaining the pluripotent state or facilitating to a specific lineage We expect that these comprehensive proteomic strategies will continue to generate new tools to study epigenetic histone PTMs and generate novel insights into the mechanism of combinatorial histone PTMs in gene regulation during diverse biological events such as cellular differentiation.