N6‐methyladenosine (m6A) methylation is the most common and abundant modification on mammalian messenger RNA (mRNA) and regulates the pluripotency of embryonic stem cells (ESCs). Research has shown that melatonin plays a fundamental role in DNA and histone modifications. However, the effect of melatonin on RNA modification is unknown. Here, for the first time, we investigated the effect of melatonin on m6A modifications in long‐term‐cultured ESCs. Pluripotency studies indicated that 10 μmol/L melatonin sufficiently maintained ESCs with stemness features over 45 passages (more than 90 days). Notably, treatment of ESCs with melatonin led to a significant decrease in the nuclear presence of m6A methyltransferase complex and decreased global m6A modification. Depletion of melatonin receptor 1 (MT1) by CRISPR/Cas9 significantly reduced the effects of melatonin on ESC pluripotency and m6A modification. Methylated RNA immunoprecipitation sequencing (MeRIP‐seq) revealed that melatonin promotes stabilization of core pluripotency factors, such as Nanog, Sox2, Klf4, and c‐Myc, by preventing m6A‐dependent mRNA decay. Using cell signaling pathway profiling systems, melatonin was shown to regulate m6A modification predominantly through the MT1‐JAK2/STAT3‐Zfp217 signal axis. This study reveals a new dimension regarding melatonin regulation of gene expression at the RNA level.

中文翻译：

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褪黑激素通过依赖褪黑激素受体的 m6A RNA 调节恢复长期培养的胚胎干细胞的多能性。

N6-甲基腺苷 (m6A) 甲基化是哺乳动物信使 RNA (mRNA) 上最常见和最丰富的修饰，可调节胚胎干细胞 (ESC) 的多能性。研究表明，褪黑激素在 DNA 和组蛋白修饰中起着重要作用。然而，褪黑激素对 RNA 修饰的影响尚不清楚。在这里，我们首次研究了褪黑激素对长期培养的 ESC 中 m6A 修饰的影响。多能性研究表明，10 μmol/L 褪黑激素足以维持具有干性特征的 ESC 超过 45 代（超过 90 天）。值得注意的是，用褪黑激素处理 ESC 导致 m6A 甲基转移酶复合物的核存在显着减少，并减少整体 m6A 修饰。CRISPR/Cas9 消耗褪黑激素受体 1 (MT1) 显着降低了褪黑激素对 ESC 多能性和 m6A 修饰的影响。甲基化 RNA 免疫沉淀测序 (MeRIP-seq) 表明，褪黑激素通过防止 m6A 依赖性 mRNA 衰减来促进核心多能性因子（如 Nanog、Sox2、Klf4 和 c-Myc）的稳定。使用细胞信号通路分析系统，显示褪黑激素主要通过 MT1-JAK2/STAT3-Zfp217 信号轴调节 m6A 修饰。这项研究揭示了关于褪黑激素在 RNA 水平上调控基因表达的新维度。通过防止 m6A 依赖性 mRNA 衰减。使用细胞信号通路分析系统，显示褪黑激素主要通过 MT1-JAK2/STAT3-Zfp217 信号轴调节 m6A 修饰。这项研究揭示了关于褪黑激素在 RNA 水平上调控基因表达的新维度。通过防止 m6A 依赖性 mRNA 衰减。使用细胞信号通路分析系统，显示褪黑激素主要通过 MT1-JAK2/STAT3-Zfp217 信号轴调节 m6A 修饰。这项研究揭示了关于褪黑激素在 RNA 水平上调控基因表达的新维度。