Li Mengyao, Li Jie, Wang Yibo, Zhao Jianxin, Yuan Aiyun, Dong Weidong, Kong Linlin, Dong Suwei, Qin Weijie, Yang Yun-Gui, Wang Xiaohui, Wu Chen, Li Jing
College of Life Sciences, Institute of Life Sciences and Green Development, Hebei University, Baoding 071002, China.
Beijing Key Laboratory of DNA Damage Response, College of Life Sciences, Capital Normal University, Beijing 100048, China.
Fundam Res. 2023 Jun 5;5(2):868-879. doi: 10.1016/j.fmre.2023.04.017. eCollection 2025 Mar.
N6-methyladenosine (mA) is the most prevalent internal RNA modification, and its regulators include writers, readers and erasers. mA is under stringent control and takes part in many biological events, but it is not known whether there is an interplay between mA and glycosylation. Here we investigated an mA reader, YTHDC1, which has been shown to be recruited to the DNA-RNA hybrid at DNA damage sites and regulate homologous recombination (HR) during DNA damage repair. We found that YTHDC1 is subject to O-linked β-N-acetylglucosamine (O-GlcNAc) modification at Ser396 upon DNA damage, which is pivotal for YTHDC1 chromatin binding and ionization radiation induced focus (IRIF) formation. RNA immunoprecipitation (RIP) and molecular dynamics (MD) simulations indicate that O-GlcNAcylation is vital for YTHDC1 to bind with mA RNA. Fluorescence recovery after photo bleaching (FRAP) analysis revealed that YTHDC1 O-GlcNAcylation is essential for DNA damage-induced YTHDC1-mA condensate formation. We further demonstrate that YTHDC1 O-GlcNAcylation promotes HR-mediated DNA damage repair and cell survival, probably through recruitment of Rad51 to the damage sites. We propose that YTHDC1 O-GlcNAcylation is instrumental for HR.
N6-甲基腺苷（mA）是最普遍的内部RNA修饰，其调控因子包括写入器、读取器和擦除器。mA受到严格调控并参与许多生物学事件，但尚不清楚mA与糖基化之间是否存在相互作用。在这里，我们研究了一种mA读取器YTHDC1，它已被证明在DNA损伤位点被招募到DNA-RNA杂交体上，并在DNA损伤修复过程中调节同源重组（HR）。我们发现，DNA损伤时，YTHDC1在Ser396位点发生O-连接的β-N-乙酰葡糖胺（O-GlcNAc）修饰，这对YTHDC1与染色质结合和电离辐射诱导灶（IRIF）形成至关重要。RNA免疫沉淀（RIP）和分子动力学（MD）模拟表明，O-GlcNAcylation对于YTHDC1与mA RNA结合至关重要。光漂白后荧光恢复（FRAP）分析显示，YTHDC1的O-GlcNAcylation对于DNA损伤诱导的YTHDC1-mA凝聚物形成至关重要。我们进一步证明，YTHDC1的O-GlcNAcylation可能通过将Rad51招募到损伤位点来促进HR介导的DNA损伤修复和细胞存活。我们提出，YTHDC1的O-GlcNAcylation对HR起作用。
