Alteration of mRNA 5-Methylcytosine Modification in Neurons After OGD/R and Potential Roles in Cell Stress Response and Apoptosis.
Alteration of mRNA 5-Methylcytosine Modification in Neurons After OGD/R and Potential Roles in Cell Stress Response and Apoptosis.
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OGD/R后神经元mRNA 5-甲基胞嘧啶修饰的变化及其在细胞应激反应和凋亡中的作用
DOI：
10.3389/fgene.2021.633681
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发表时间：
2021
影响因子：
3.7
通讯作者：
Feng S
中科院分区：
文献类型：
--
作者：
Jian H;Zhang C;Qi Z;Li X;Lou Y;Kang Y;Deng W;Lv Y;Wang C;Wang W;Shang S;Hou M;Zhou H;Feng S
表观遗传修饰在中枢神经系统疾病中起重要作用。作为一种广泛存在的RNA转录后修饰，m5 C修饰在脑缺血再灌注损伤（IRI）中的作用尚不清楚。在这里，我们成功地构建了一个神经元氧-葡萄糖剥夺/复氧（OGD/R）模型，并使用RNA-BS-seq获得了全转录组m5 C谱的概述。我们发现神经元m5 C修饰的分布是高度保守的，显著富集在富含CG的区域，并集中在mRNA翻译起始区域。OGD/R后，m5 C基因的修饰水平增加，甲基化基因的数目减少。m5 C位点与大多数RNA结合蛋白结合位点的重叠量显著增加，RBM 3结合蛋白除外。此外，神经元中的高甲基化基因在病理过程中显著富集，蛋白质-蛋白质相互作用网络鉴定的中枢高甲基化基因RPL 8和RPS 9与脑损伤显著相关。此外，与高甲基化修饰的上调的转录物中富集的过程中参与应激反应和细胞凋亡的调节，这些过程中未发现低甲基化的转录物。最后，我们用TUNEL和Western blot检测证实了OGD/R诱导的神经元凋亡。我们的研究发现了与神经元缺血再灌注相关的新m5 C mRNA，为进一步研究RNA甲基化在脑IRI中的作用提供了有价值的观点。
Epigenetic modifications play an important role in central nervous system disorders. As a widespread posttranscriptional RNA modification, the role of the m5C modification in cerebral ischemia-reperfusion injury (IRI) remains poorly defined. Here, we successfully constructed a neuronal oxygen-glucose deprivation/reoxygenation (OGD/R) model and obtained an overview of the transcriptome-wide m5C profiles using RNA-BS-seq. We discovered that the distribution of neuronal m5C modifications was highly conserved, significantly enriched in CG-rich regions and concentrated in the mRNA translation initiation regions. After OGD/R, modification level of m5C increased, whereas the number of methylated mRNA genes decreased. The amount of overlap of m5C sites with the binding sites of most RNA-binding proteins increased significantly, except for that of the RBM3-binding protein. Moreover, hypermethylated genes in neurons were significantly enriched in pathological processes, and the hub hypermethylated genes RPL8 and RPS9 identified by the protein-protein interaction network were significantly related to cerebral injury. Furthermore, the upregulated transcripts with hypermethylated modification were enriched in the processes involved in response to stress and regulation of apoptosis, and these processes were not identified in hypomethylated transcripts. In final, we verified that OGD/R induced neuronal apoptosis in vitro using TUNEL and western blot assays. Our study identified novel m5C mRNAs associated with ischemia-reperfusion in neurons, providing valuable perspectives for future studies on the role of the RNA methylation in cerebral IRI.