Understanding the Role of Combinatorial Histone PTM Patterns

描述（由申请人提供）：组蛋白的翻译后修饰（PTM）是真核生物中调节染色质结构和基因表达模式的主要表观遗传机制。除了它们在标准生理学中的重要作用之外，组蛋白PTM信号传导模式的破坏已经被认为是各种人类疾病如癌症中的重要的潜在致病因素。由于染色质生物学领域中的大多数组蛋白PTM工作是使用位点特异性抗体完成的，因此在同一分子上共发生的组合组蛋白PTM的定量测量尚未得到满足。我们的目标包括继续发展基于质谱的蛋白质组学和生物信息学方法，用于定量询问组合组蛋白PTM模式，并应用这些方法研究健康相关生物学研究关键领域背后的组蛋白介导的表观遗传学机制。在这里，我们将专门应用我们的方法来研究人类胚胎干细胞分化过程中的表观遗传组蛋白PTM信号。我们的具体目标有三个，包括鉴定干细胞分化过程中变化的组蛋白PTM，表征翻译这些PTM模式的组合组蛋白PTM结合蛋白复合物，并确定这些组合PTM在维持多能状态或促进特定谱系中的作用。我们期望这些全面的蛋白质组学策略将继续产生新的工具来研究表观遗传组蛋白PTM，产生新的见解组合组蛋白PTM在基因调控的机制，在不同的生物事件，如细胞分化。

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.