FTO regulates the DNA damage response via effects on cell-cycle progression
FTO regulates the DNA damage response via effects on cell-cycle progression
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DOI：
10.1016/j.mrgentox.2023.503608
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发表时间：
2023-03-01
影响因子：
1.9
通讯作者：
Luan, Yang
中科院分区：
文献类型：
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作者：
Liu, Weiying;Yasui, Manabu;Luan, Yang
脂肪量和肥胖相关蛋白FTO是N6-甲基腺苷的“橡皮擦”，N6-甲基腺苷是最丰富的mRNA修饰。FTO在肿瘤发生中起重要作用。然而，其活动尚未完全阐明，其可能参与DNA损伤-肿瘤发生的早期驱动事件-仍然缺乏特征。在这里，我们研究了FTO在DNA损伤反应（DDR）中的作用及其潜在机制。我们证明，FTO响应各种DNA损伤刺激。FTO在暴露于前诱变剂马兜铃酸I和苯并[a]芘的小鼠中过表达。通过CRISPR/Cas9敲除TK 6细胞中的FTO基因，增加了DNA损伤刺激物诱导的遗传毒性（微核和TK突变测定）。顺铂和二环氧丁烷诱导的微核频率以及甲磺酸甲酯和硫唑嘌呤诱导的TK突变频率在FTO KO细胞中也较高。我们调查了FTO在复员方案中的潜在作用。RNA测序和富集分析显示，FTO缺失破坏了p38 MAPK通路，并抑制了顺铂（DNA链内交联）治疗后核苷酸切除修复和细胞周期相关通路的激活。这些作用通过蛋白质印迹和qRT-PCR证实。FTO缺失损害顺铂和二环氧丁烷处理后细胞周期停滞在G2/M期（流式细胞术分析）。我们的研究结果表明，FTO参与了DDR的几个方面，至少部分通过损害细胞周期进程发挥作用。
The fat mass and obesity-associated protein FTO is an "eraser" of N6-methyladenosine, the most abundant mRNA modification. FTO plays important roles in tumorigenesis. However, its activities have not been fully elucidated and its possible involvement in DNA damage - the early driving event in tumorigenesis - remains poorly characterized. Here, we have investigated the role of FTO in the DNA damage response (DDR) and its underlying mechanisms. We demonstrate that FTO responds to various DNA damage stimuli. FTO is overexpressed in mice following exposure to the promutagens aristolochic acid I and benzo[a]pyrene. Knockout of the FTO gene in TK6 cells, via CRISPR/Cas9, increased genotoxicity induced by DNA damage stimuli (micronucleus and TK mutation assays). Cisplatin- and diepoxybutane-induced micronucleus frequencies and methyl methanesulfonate- and azathioprine-induced TK mutant frequencies were also higher in FTO KO cells. We investigated the potential roles of FTO in DDR. RNA sequencing and enrichment analysis revealed that FTO deletion disrupted the p38 MAPK pathway and inhibited the activation of nucleotide excision repair and cell-cycle-related pathways following cisplatin (DNA intrastrand cross-links) treatment. These effects were confirmed by western blotting and qRT-PCR. FTO deletion impaired cell-cycle arrest at the G2/M phase following cisplatin and diepoxybutane treatment (flow cytometry analysis). Our findings demonstrated that FTO is involved in several aspects of DDR, acting, at least in part, by impairing cell cycle progression.