RNA N⁶-methyladenosine (m⁶A) modification plays important roles in multiple aspects of RNA regulation. m⁶A is installed cotranscriptionally by the METTL3/14 complex, but its direct roles in RNA processing remain unclear. Here, we investigate the presence of m⁶A in nascent RNA of mouse embryonic stem cells. We find that around 10% of m⁶A peaks are located in alternative introns/exons, often close to 5′ splice sites. m⁶A peaks significantly overlap with RBM15 RNA binding sites and the histone modification H3K36me3. Acute depletion of METTL3 disrupts inclusion of alternative introns/exons in the nascent transcriptome, particularly at 5′ splice sites that are proximal to m⁶A peaks. For terminal or variable-length exons, m⁶A peaks are generally located on or immediately downstream from a 5′ splice site that is suppressed in the presence of m⁶A and upstream of a 5′ splice site that is promoted in the presence of m⁶A. Genes with the most immediate effects on splicing include several components of the m⁶A pathway, suggesting an autoregulatory function. Collectively, our findings demonstrate crosstalk between the m⁶A machinery and the regulation of RNA splicing.

中文翻译：

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METTL3 的急性耗竭暗示 N6-甲基腺苷在新生转录组中的替代内含子/外显子包含中

RNA N ⁶ -甲基腺苷 (m ⁶ A) 修饰在 RNA 调控的多个方面发挥着重要作用。m ⁶ A 由 METTL3/14 复合体以共转录方式安装，但其在 RNA 加工中的直接作用仍不清楚。在这里，我们研究了小鼠胚胎干细胞的新生 RNA 中m ^{6 A 的存在。}我们发现大约 10% 的 m ⁶ A 峰位于替代内含子/外显子中，通常靠近 5' 剪接位点。m ⁶ A 峰与 RBM15 RNA 结合位点和组蛋白修饰 H3K36me3 显着重叠。METTL3 的急性消耗破坏了新生转录组中替代内含子/外显子的包含，特别是在靠近 m 的 5' 剪接位点^6个A峰。对于末端或可变长度外显子，m ⁶ A 峰通常位于存在 m 6 A 时被抑制的 5' 剪接位点上或紧邻下游，以及在存在 m ⁶ A 时被促进的 5' 剪接位点上游。 m ⁶ A. 对剪接产生最直接影响的基因包括 m ⁶ A 途径的几个组成部分，表明具有自动调节功能。总的来说，我们的研究结果证明了 m ⁶ A 机器与 RNA 剪接调节之间的串扰。