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首页> 外文期刊>Cell biology international. >Nanog RNA-binding proteins YBX1 and ILF3 affect pluripotency of embryonic stem cells
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Nanog RNA-binding proteins YBX1 and ILF3 affect pluripotency of embryonic stem cells

机译：Nanog RNA结合蛋白YBX1和ILF3影响胚胎干细胞的多能性

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Nanog is a well-known transcription factor that plays a fundamental role in stem cell self-renewal and the maintenance of their pluripotent cell identity. There remains a large data gap with respect to the spectrum of the key pluripotency transcription factors' interaction partners. Limited information is available concerning Nanog-associated RNA-binding proteins (RBPs), and the intrinsic protein-RNA interactions characteristic of the regulatory activities of Nanog. Herein, we used an improved affinity protocol to purify Nanog-interacting RBPs from mouse embryonic stem cells (ESCs), and 49 RBPs of Nanog were identified. Among them, the interaction of YBX1 and ILF3 with Nanog mRNA was further confirmed by in vitro assays, such as Western blot, RNA immunoprecipitation (RIP), and ex vivo methods, such as immunofluorescence staining and fluorescent in situ hybridization (FISH), MS2 in vivo biotin-tagged RNA affinity purification (MS2-BioTRAP). Interestingly, RNAi studies revealed that YBX1 and ILF3 positively affected the expression of Nanog and other pluripotency-related genes. Particularly, downregulation of YBX1 or ILF3 resulted in high expression of mesoderm markers. Thus, a reduction in the expression of YBX1 and ILF3 controls the expression of pluripotency-related genes in ESCs, suggesting their roles in further regulation of the pluripotent state of ESCs.
机译：Nanog是一种众所周知的转录因子，在干细胞自我更新和维持其多能细胞特性中起着基本作用。关于关键多能转录因子的相互作用伙伴的光谱，仍然存在较大的数据缺口。关于Nanog相关的RNA结合蛋白（RBP），以及Nanog调节活性的内在蛋白质-RNA相互作用特征，目前尚缺乏有限的信息。在这里，我们使用改进的亲和协议从小鼠胚胎干细胞（ESC）中纯化与Nanog相互作用的RBP，并鉴定出49个Nanog的RBP。其中，YBX1和ILF3与Nanog mRNA的相互作用已通过体外测定（如Western blot，RNA免疫沉淀（RIP））和离体方法（如免疫荧光染色和荧光原位杂交（FISH），MS2）得到进一步证实。体内生物素标记的RNA亲和纯化（MS2-BioTRAP）。有趣的是，RNAi研究表明YBX1和ILF3对Nanog和其他多能性相关基因的表达有积极影响。特别是，YBX1或ILF3的下调导致中胚层标记物的高表达。因此，YBX1和ILF3表达的减少控制着多能性相关基因在ESC中的表达，表明它们在进一步调节ESC多能性状态中的作用。

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