Interdisciplinary Program in Agricultural Genomics, Department of Plant Science, Plant Genomics and Breeding Institute, and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, 08826, Korea.
School of Biological Sciences, Seoul National University, Seoul, 08826, Korea.
Sci Rep. 2017 Aug 22;7(1):9160. doi: 10.1038/s41598-017-08827-9.
DNA methylation is a prominent epigenetic modification in plants and animals regulated by similar mechanisms but the process of DNA demethylation is profoundly different. Unlike vertebrates that require a series of enzymatic conversions of 5-methylcytosine (5mC) into other bases for DNA demethylation, plants utilize the DEMETER (DME) family of 5mC DNA glycosylases to catalyze a direct removal of 5mC from DNA. Here we introduced Arabidopsis DME into human HEK-293T cells to allow direct 5mC excision, and observed that direct DNA demethylation activity was successfully implemented by DME expression. In addition, DME induced diverse cellular responses such as cell proliferation inhibition, cell cycle dysregulation and S phase arrest. Microarray and methylome analyses revealed that DME upregulated a number of genes including cell cycle components, heat shock proteins, and notably, various interferon-stimulated genes. Moreover, DME-mediated DNA demethylation activated endogenous repeat elements, which are likely to form dsRNAs as viral mimics and eventually trigger interferon cascades to establish the antiviral state. This work demonstrates that plant DNA demethylase catalyzes DNA demethylation with a bypass of initial base conversion steps, and the interferon signaling plays a pivotal role to alleviate genotoxic stresses associated with DME-induced DNA demethylation in mammalian cells.
DNA 甲基化是动植物中一种重要的表观遗传修饰,受相似的机制调控,但 DNA 去甲基化的过程却大不相同。与需要一系列酶促转化将 5-甲基胞嘧啶(5mC)转化为其他碱基才能进行 DNA 去甲基化的脊椎动物不同,植物利用 DEMETER(DME)家族的 5mC DNA 糖苷酶直接从 DNA 中切除 5mC。在这里,我们将拟南芥 DME 引入人 HEK-293T 细胞中,以允许直接切除 5mC,并观察到 DME 表达成功实现了直接 DNA 去甲基化活性。此外,DME 诱导了多种细胞反应,如细胞增殖抑制、细胞周期失调和 S 期阻滞。微阵列和甲基组分析表明,DME 上调了许多基因,包括细胞周期成分、热休克蛋白,特别是各种干扰素刺激基因。此外,DME 介导的 DNA 去甲基化激活了内源性重复元件,这些元件可能形成 dsRNA,作为病毒模拟物,并最终触发干扰素级联反应,建立抗病毒状态。这项工作表明,植物 DNA 去甲基酶催化 DNA 去甲基化,绕过初始碱基转换步骤,干扰素信号通路在减轻与 DME 诱导的哺乳动物细胞 DNA 去甲基化相关的遗传毒性应激方面发挥关键作用。