Polyglutamine-expanded ataxin3 alter specific gene expressions through changing DNA methylation status in SCA3/MJD.
Polyglutamine-expanded ataxin3 alter specific gene expressions through changing DNA methylation status in SCA3/MJD.
复制标题
聚谷氨酰胺扩增的ataxin3通过改变SCA3/MJD中的DNA甲基化状态来改变特定基因表达
DOI：
10.18632/aging.202331
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发表时间：
2020-12-19
期刊：
影响因子：
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通讯作者：
Jiang H
中科院分区：
文献类型：
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作者：
Ding D;Wang C;Chen Z;Xia K;Tang B;Qiu R;Jiang H
DNA甲基化最近被认为与多聚谷氨酰胺(PolyQ)病的转录失调和神经元功能障碍有关。本研究旨在确定ATXN3中(CAG)n的扩展是否扰乱DNA甲基化状态并影响基因表达。我们使用简化代表性亚硫酸氢盐测序(RRBS)分析了整个基因组的DNA甲基化，并使用甲基靶测序证实了结果。用BSP、qRT-PCR和Western印迹检测特定基因的DNA甲基化、转录和翻译水平的动态变化。总共在SCA3/MJD和WT小鼠小脑中鉴定出135个差异甲基化区域(DMR)。KEGG分析显示差异甲基化基因涉及氨基酸代谢、Hedgehog信号通路、甲状腺癌、肿瘤发生等途径。我们把重点放在与基因表达直接相关的DMR上。在此基础上，我们进一步评估了7个基因，包括13个DMR，用于DNA甲基化验证和基因表达。我们发现EN1和Nkx2-1的DMR的甲基化状态与它们的转录和翻译水平呈负相关，在SCA3/MJD小鼠运动障碍之前，DMR的DNA甲基化状态和相应的转录发生了变化。这些结果揭示了新的DNA甲基化调控基因EN1和Nkx2-1，这可能有助于理解SCA3/MJD的发病机制。
DNA methylation has recently been linked to transcriptional dysregulation and neuronal dysfunction in polyglutamine (polyQ) disease. This study aims to determine whether (CAG)n expansion in ATXN3 perturbs DNA methylation status and affects gene expression. We analyzed DNA methylation throughout the genome using reduced representation bisulfite sequencing (RRBS) and confirmed the results using MethylTarget sequencing. Dynamic changes in DNA methylation, transcriptional and translational levels of specific genes were detected using BSP, qRT-PCR and western blot. In total, 135 differentially methylated regions (DMRs) were identified between SCA3/MJD and WT mouse cerebellum. KEGG analysis revealed differentially methylated genes involved in amino acid metabolism, Hedgehog signaling pathway, thyroid cancer, tumorigenesis and other pathways. We focused on DMRs that were directly associated with gene expression. On this basis, we further assessed 7 genes, including 13 DMRs, for DNA methylation validation and gene expression. We found that the methylation status of the DMRs of En1 and Nkx2-1 was negatively associated with their transcriptional and translational levels and that alteration of the DNA methylation status of DMRs and the corresponding transcription occurred before dyskinesia in SCA3/MJD mice. These results revealed novel DNA methylation-regulated genes, En1 and Nkx2-1, which may be useful for understanding the pathogenesis of SCA3/MJD.