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TET1 Interacts Directly with NANOG via Independent Domains Containing Hydrophobic and Aromatic Residues
Journal of Molecular Biology ( IF 5.6 ) Pub Date : 2020-10-13 , DOI: 10.1016/j.jmb.2020.10.008
Raphaël Pantier , Nicholas Mullin , Elisa Hall-Ponsele , Ian Chambers

The DNA demethylase TET1 is highly expressed in embryonic stem cells and is important both for lineage commitment, and reprogramming to naïve pluripotency. TET1 interacts with the pluripotency transcription factor NANOG which may contribute to its biological activity in pluripotent cells. However, how TET1 interacts with other proteins is largely unknown. Here, we characterise the physical interaction between TET1 and NANOG using embryonic stem cells and bacterial expression systems. TET1 and NANOG interact through multiple binding sites that act independently. Critically, mutating conserved hydrophobic and aromatic residues within TET1 and NANOG abolishes the interaction. On chromatin, NANOG is predominantly localised at ESC enhancers. While TET1 binds to CpG dinucleotides in promoters using its CXXC domain, TET1 also binds to enhancers, though the mechanism involved is unknown. Comparative ChIP-seq analysis identifies genomic loci bound by both TET1 and NANOG, that correspond predominantly to pluripotency enhancers. Importantly, around half of NANOG transcriptional target genes are associated with TET1-NANOG co-bound sites. These results indicate a mechanism by which TET1 protein may be targeted to specific sites of action at enhancers by direct interaction with a transcription factor.



中文翻译:

TET1通过包含疏水和芳香残基的独立域与NANOG直接相互作用

DNA脱甲基酶TET1在胚胎干细胞中高度表达,对于谱系承诺和重编程以达到幼稚的多能性都很重要。TET1与多能转录因子NANOG相互作用,这可能有助于其在多能细胞中的生物学活性。但是,TET1如何与其他蛋白质相互作用的方法在很大程度上尚不清楚。在这里,我们表征利用胚胎干细胞和细菌表达系统的TET1和NANOG之间的物理相互作用。TET1和NANOG通过多个独立起作用的结合位点相互作用。至关重要的是,TET1和NANOG中突变的保守疏水和芳香残基会消除相互作用。在染色质上,NANOG主要位于ESC增强子上。TET1使用其CXXC域与启动子中的CpG二核苷酸结合,而TET1也与增强子结合,尽管涉及的机制尚不清楚。比较ChIP-seq分析可确定受TET1和NANOG结合的基因组位点,它们主要对应于多能性增强子。重要的是,大约一半的NANOG转录靶基因与TET1-NANOG共同结合位点相关。这些结果表明一种机制,通过该机制,TET1蛋白可通过与转录因子的直接相互作用靶向增强子的特定作用位点。

更新日期:2020-11-23
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