Vlachogiannis Georgios, Niederhuth Chad E, Tuna Salih, Stathopoulou Athanasia, Viiri Keijo, de Rooij Dirk G, Jenner Richard G, Schmitz Robert J, Ooi Steen K T
Epigenetic Signaling Group, Department of Cancer Biology, UCL Cancer Institute, Paul O'Gorman Building, 72 Huntley Street, London WC1E 6BT, UK.
Department of Genetics, University of Georgia, 120 East Green Street, Athens, GA 30602, USA.
Cell Rep. 2015 Feb 17;10(6):944-956. doi: 10.1016/j.celrep.2015.01.021. Epub 2015 Feb 13.
A critical aspect of mammalian gametogenesis is the reprogramming of genomic DNA methylation. The catalytically inactive adaptor Dnmt3L is essential to ensuring this occurs correctly, but the mechanism by which it functions is unclear. Using gene targeting to engineer a single-amino-acid mutation, we show that the Dnmt3L histone H3 binding domain (ADD) is necessary for spermatogenesis. Genome-wide single-base-resolution DNA methylome analysis of mutant germ cells revealed overall reductions in CG methylation at repetitive sequences and non-promoter CpG islands. Strikingly, we also observe an even more severe loss of non-CG methylation, suggesting an unexpected role for the ADD in this process. These epigenetic deficiencies were coupled with defects in spermatogonia, with mutant cells displaying marked changes in gene expression and reactivation of retrotransposons. Our results demonstrate that the Dnmt3L ADD is necessary for Dnmt3L function and full reproductive fitness.
哺乳动物配子发生的一个关键方面是基因组DNA甲基化的重编程。催化无活性的衔接蛋白Dnmt3L对于确保这一过程正确发生至关重要，但其发挥作用的机制尚不清楚。通过基因靶向工程制造一个单氨基酸突变，我们发现Dnmt3L组蛋白H3结合结构域（ADD）对精子发生是必需的。对突变生殖细胞进行全基因组单碱基分辨率DNA甲基化组分析发现，重复序列和非启动子CpG岛处的CG甲基化总体减少。引人注目的是，我们还观察到非CG甲基化的损失更为严重，这表明ADD在这一过程中具有意想不到的作用。这些表观遗传缺陷与精原细胞的缺陷相关，突变细胞在基因表达和逆转录转座子的重新激活方面表现出明显变化。我们的结果表明，Dnmt3L ADD对于Dnmt3L功能和完全生殖适应性是必需的。
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