Hao Yi, Li Xiang, Qin Ke, Shi Yujie, He Yanwen, Zhang Che, Cheng Bo, Zhang Xiwen, Hu Guangyu, Liang Shuyu, Tang Qi, Chen Xing
College of Chemistry and Molecular Engineering, Beijing National Laboratory for Molecular Sciences, Peking-Tsinghua Center for Life Sciences, Synthetic and Functional Biomolecules Center, Key Laboratory of Bioorganic Chemistry and Molecular Engineering of Ministry of Education, Peking University, Beijing, 100871, China.
Angew Chem Int Ed Engl. 2023 Apr 17;62(17):e202300500. doi: 10.1002/anie.202300500. Epub 2023 Mar 20.
Self-renewal and differentiation of embryonic stem cells (ESCs) are influenced by protein O-linked β-N-acetylglucosamine (O-GlcNAc) modification, but the underlying mechanism remains incompletely understood. Herein, we report the identification of 979 O-GlcNAcylated proteins and 1340 modification sites in mouse ESCs (mESCs) by using a chemoproteomics method. In addition to OCT4 and SOX2, the third core pluripotency transcription factor (PTF) NANOG was found to be modified and functionally regulated by O-GlcNAc. Upon differentiation along the neuronal lineage, the O-GlcNAc stoichiometry at 123 sites of 83 proteins-several of which were PTFs-was found to decline. Transcriptomic profiling reveals 2456 differentially expressed genes responsive to OGT inhibition during differentiation, of which 901 are target genes of core PTFs. By acting on the core PTF network, suppression of O-GlcNAcylation upregulates neuron-related genes, thus contributing to mESC fate determination.
胚胎干细胞（ESC）的自我更新和分化受蛋白质O-连接β-N-乙酰葡糖胺（O-GlcNAc）修饰的影响，但其潜在机制仍未完全阐明。在此，我们报告了通过化学蛋白质组学方法在小鼠胚胎干细胞（mESC）中鉴定出979种O-GlcNAc化蛋白和1340个修饰位点。除了OCT4和SOX2外，还发现第三个核心多能性转录因子（PTF）NANOG被O-GlcNAc修饰并受到功能调控。在沿神经谱系分化时，发现83种蛋白质（其中几种是PTF）的123个位点上的O-GlcNAc化学计量下降。转录组分析揭示了2456个在分化过程中对OGT抑制有反应的差异表达基因，其中901个是核心PTF的靶基因。通过作用于核心PTF网络，抑制O-GlcNAc化会上调神经元相关基因，从而有助于mESC命运的决定。
