Structure and Functionof Nanog in Stem Cell Pluripotency

项目摘要 干细胞多能性中的NANOG结构和功能 NANOG指导获得干细胞多能性。在我们的研究中， 分子理解NANOG如何协调这一过程。此外， 事实上，NANOG（特别是人类NANOG）一直是一个具有挑战性的系统， 以.研究人员成功地将创新的荧光技术应用于 表征NANOG的关键结构和功能特性。NANOG本质上是一种 无序蛋白（IDP），具有对NANOG自组装和对 在多能性特异性区域中建立接触。NANOG是一个“分子枢纽”，据报道， 与>100个转录因子（TF）和其他蛋白质相互作用，形成合作中心 多能性基因的凝聚和协调激活以及分化的抑制 基因.研究人员将调查和解卷积NANOG剂量敏感性如何发挥作用， 在染色质成环和转录因子、共激活因子和表观遗传调节因子的募集中的作用。他们 将使用标准和创新的结构，分子， 基因组学和细胞生物学方法。第一个目标将阐明NANOG的剂量敏感性 通过评估染色质重组和募集建立多能性的机制 使用Hi-C 3.0和单分子荧光技术的辅因子。第二个目标将 探测分子元件（即，OCT 4-SOX 2-NANOG三联体协同性和KLF 4 生物分子缩合）参与激活NANOG表达。此外，目标将 研究NANOG自动阻遏的分子基础。最后一个目标将侧重于抑制 的NANOG稳定化和寡聚化与显性负NANOG为基础的肽。 这些研究将为NANOG如何协调建立提供前所未有的见解 干细胞多能性和逆转发育性衰老。

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.