Structure and Functionof Nanog in Stem Cell Pluripotency

项目摘要 干细胞多能性中的Nanog结构与功能 Nanog指导人们获得干细胞的多能性。我们有相当大的差距 对NANOG如何协调这一过程的分子理解。这一点还会因为 NANOG(尤其是人类NANOG)一直是一个具有挑战性的系统 和.。研究人员成功地将创新的荧光技术应用于 描述NANOG的关键结构和功能特性。Nanog是一种本质上的 无序蛋白(IDP)，其具有一个Pron样结构域，对NANOG自组装和 在多潜能特定地区建立联系。Nanog是一个“分子枢纽”，据报道 与&gt；100转录因子(TF)和其他蛋白质相互作用形成协同中心 凝聚和协调多能基因的激活和分化的抑制 基因。调查人员将调查和揭开NANOG剂量敏感性是如何发挥作用的 在染色质循环和募集转录因子、共激活因子和表观遗传调节因子中的作用。他们 将使用标准和创新的结构、分子、 基因组和细胞生物学方法。第一个目标是阐明NANOG的剂量敏感性 通过评估染色质重组和招募来建立多能性的机制 使用Hi-C 3.0和单分子荧光技术对辅因子进行分析。第二个目标是 探索分子元素(即OCT4-SOX2-NANOG三联体协同作用和KLF4 生物分子缩合)参与激活NANOG的表达。此外，目标将是 研究NANOG自阻抗的分子基础。最后一个目标将集中在抑制上 NANOG稳定和以负性NANOG为主的多肽的齐聚。 这些研究将为NANOG如何协调建立提供前所未有的见解 干细胞的多能性和逆转发育衰老。

Project Summary NANOG Structure and Function in Stem Cell Pluripotency NANOG directs access to stem cell pluripotency. There are considerable gaps in our molecular understanding of how NANOG coordinates this process. This is also compounded by the fact that NANOG (in particular, human NANOG) has been a challenging system to work with. The investigators have successfully applied innovative fluorescence techniques to characterize NANOG’s key structure and functional properties. NANOG is an intrinsically disordered protein (IDP), with a prion-like domain critical for NANOG self-assembly and for establishing contacts in pluripotency specific regions. NANOG is a ‘molecular hub’, reported to interact with >100 transcription factors (TFs) and other proteins to form cooperative hub condensates and coordinate activation of pluripotency genes and repression of differentiation genes. The investigators will investigate and deconvolute how NANOG dose-sensitivity play a role in chromatin looping and recruitment of TFs, co-activators, and epigenetic regulators. They will use an intensive holistic combination of standard and innovative structural, molecular, genomic and cell biology approaches. The first aim will elucidate NANOG’s dose-sensitivity mechanism in establishing pluripotency by assessing chromatin reorganization and recruitment of cofactors using Hi-C 3.0 and single molecule fluorescence techniques. The second aim will probe the molecular elements (i.e., OCT4-SOX2-NANOG triad cooperativity and KLF4 biomolecular condensation) involved in activating NANOG expression. In addition, the aim will investigate the molecular basis for NANOG autorepression. The last aim will focus on inhibition of NANOG stabilization and oligomerization with dominant negative NANOG-based peptides. These studies will provide unprecedented insights into how NANOG orchestrates establishment of stem cell pluripotency and reversal of developmental aging.