Next-generation unnatural monosaccharides reveal that ESRRB O-GlcNAcylation regulates pluripotency of mouse embryonic stem cells
Next-generation unnatural monosaccharides reveal that ESRRB O-GlcNAcylation regulates pluripotency of mouse embryonic stem cells
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DOI：
10.1038/s41467-019-11942-y
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发表时间：
2019-09-06
影响因子：
16.6
通讯作者：
Chen, Xing
中科院分区：
文献类型：
--
作者：
Hao, Yi;Fan, Xinqi;Chen, Xing
非天然单糖，如叠氮糖，可以代谢结合到细胞聚糖中，目前被用作聚糖成像和糖蛋白组学分析的主要工具。作为增强膜通透性和细胞摄取的一种常见做法，非天然糖被乙酰化，然而，这可能会引起一个长期被忽视的副反应，非酶s -糖基化。在此，我们开发了1,3-二酯化n -叠氮酰基半乳糖胺（GalNAz）作为下一代代谢聚糖标记的化学报告。1,3-二o -乙酰化的GalNAz （1,3- ac2galnaz）和1,3-二o -丙酰化的GalNAz （1,3- pr (2)GalNAz）在不需要人工s -糖基化的情况下都能高效地标记蛋白质的o - glcn酰化。利用1,3- pr (2)GalNAz在小鼠胚胎干细胞（mESCs）中的应用，我们发现ESRRB是多能性的关键转录因子，是一种o - glcn酰化蛋白。我们发现ESRRB o - glcn酰化对mESC自我更新和多能性很重要。机制上，ESRRB被O-GlcNAc转移酶在25号丝氨酸上进行O-GlcNAc酰化，这稳定了ESRRB，促进了其转录活性，并促进了其与两个主要多能性调节因子OCT4和NANOG的相互作用。
Unnatural monosaccharides such as azidosugars that can be metabolically incorporated into cellular glycans are currently used as a major tool for glycan imaging and glycoproteomic profiling. As a common practice to enhance membrane permeability and cellular uptake, the unnatural sugars are per-O-acetylated, which, however, can induce a long-overlooked side reaction, non-enzymatic S-glycosylation. Herein, we develop 1,3-di-esterified N-azidoacetylgalactosamine (GalNAz) as next-generation chemical reporters for metabolic glycan labeling. Both 1,3-di-O-acetylated GalNAz (1,3-Ac2GalNAz) and 1,3-di-O-propionylated GalNAz (1,3-Pr(2)GalNAz) exhibit high efficiency for labeling protein O-GlcNAcylation with no artificial S-glycosylation. Applying 1,3-Pr(2)GalNAz in mouse embryonic stem cells (mESCs), we identify ESRRB, a critical transcription factor for pluripotency, as an O-GlcNAcylated protein. We show that ESRRB O-GlcNAcylation is important for mESC self-renewal and pluripotency. Mechanistically, ESRRB is O-GlcNAcylated by O-GlcNAc transferase at serine 25, which stabilizes ESRRB, promotes its transcription activity and facilitates its interactions with two master pluripotency regulators, OCT4 and NANOG.