No evidence for AID/MBD4-coupled DNA demethylation in zebrafish embryos.
No evidence for AID/MBD4-coupled DNA demethylation in zebrafish embryos.
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DOI：
10.1371/journal.pone.0114816
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Kikuchi Y
中科院分区：
文献类型：
--
作者：
Shimoda N;Hirose K;Kaneto R;Izawa T;Yokoi H;Hashimoto N;Kikuchi Y
在后生动物中，负责活性DNA去甲基化的机制仍然难以捉摸。先前的一项利用斑马鱼胚胎的研究提供了一种有效的活性去甲基化机制，其中涉及三种蛋白质，AID， MBD4和GADD45。我们最近在斑马鱼中发现了年龄依赖性的DNA低甲基化，这促使我们研究AID和MBD4是否可能参与了这种现象。然而，出乎意料的是，我们发现之前研究的大部分结果都是不可重复的。首先，将甲基化的DNA片段注射到斑马鱼卵中，既不影响基因组DNA的甲基化，也不影响注射的甲基化DNA本身，也不影响检测的几个位点或aid的表达水平，aid已被证明在去甲基化中发挥作用。其次，在注射抗aid和mbd4的反义morpholino寡核苷酸后，在某些CpG岛未观察到异常甲基化。此外，我们证明了斑马鱼MBD4 cDNA缺乏甲基- cpg结合域的编码区，这被认为是引导目标区域所必需的。综上所述，我们得出结论，目前没有证据支持AID和MBD4在斑马鱼胚胎活性去甲基化中的作用。
The mechanisms responsible for active DNA demethylation remain elusive in Metazoa. A previous study that utilized zebrafish embryos provided a potent mechanism for active demethylation in which three proteins, AID, MBD4, and GADD45 are involved. We recently found age-dependent DNA hypomethylation in zebrafish, and it prompted us to examine if AID and MBD4 could be involved in the phenomenon. Unexpectedly, however, we found that most of the findings in the previous study were not reproducible. First, the injection of a methylated DNA fragment into zebrafish eggs did not affect either the methylation of genomic DNA, injected methylated DNA itself, or several loci tested or the expression level of aid, which has been shown to play a role in demethylation. Second, aberrant methylation was not observed at certain CpG islands following the injection of antisense morpholino oligonucleotides against aid and mbd4. Furthermore, we demonstrated that zebrafish MBD4 cDNA lacked a coding region for the methyl-CpG binding domain, which was assumed to be necessary for guidance to target regions. Taken together, we concluded that there is currently no evidence to support the proposed roles of AID and MBD4 in active demethylation in zebrafish embryos.
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影响因子：
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Ekker, M