Chemoproteomic and Transcriptomic Analysis Reveals that O-GlcNAc Regulates Mouse Embryonic Stem Cell Fate through the Pluripotency Network.
Yi HaoXiang LiKe QinYujie ShiYanwen HeChe ZhangBo ChengXiwen ZhangGuangyu HuShuyu LiangQi TangXing ChenPublished in: Angewandte Chemie (International ed. in English) (2023)
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.
Keyphrases
- embryonic stem cells
- cell fate
- transcription factor
- single cell
- bioinformatics analysis
- genome wide identification
- genome wide
- rna seq
- stem cells
- gene expression
- solid phase extraction
- dna binding
- blood brain barrier
- high resolution
- dna methylation
- mass spectrometry
- cerebral ischemia
- cancer stem cells
- liquid chromatography