Live-cell analysis of DNA methylation during sexual reproduction in Arabidopsis reveals context and sex-specific dynamics controlled by noncanonical RdDM.
Live-cell analysis of DNA methylation during sexual reproduction in Arabidopsis reveals context and sex-specific dynamics controlled by noncanonical RdDM.
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DOI：
10.1101/gad.289397.116
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发表时间：
2017-01-01
影响因子：
10.5
通讯作者：
Grimanelli D
中科院分区：
文献类型：
--
作者：
Ingouff M;Selles B;Michaud C;Vu TM;Berger F;Schorn AJ;Autran D;Van Durme M;Nowack MK;Martienssen RA;Grimanelli D
在这个资源/方法中，Ingouff等人开发了一种新的方法来研究植物中的DNA甲基化重编程。使用拟南芥，作者开发了传感器，以单细胞分辨率报告CG和非CG甲基化。胞嘧啶甲基化是许多生物体中的关键表观遗传标记，对于转录控制和基因组完整性都很重要。虽然在体细胞生长期间相对稳定，但DNA甲基化在哺乳动物生殖期间在全基因组范围内重新编程。重编程对于合子全能性和防止表位突变的跨代遗传是必不可少的。然而，植物中DNA甲基化重编程的程度仍不清楚。在这里，我们开发了传感器报告与单细胞分辨率CG和非CG甲基化拟南芥。在繁殖过程中的现场成像揭示了不同的和性别特异性的动态这两种情况。我们发现，卵细胞中CHH甲基化依赖于结构域重排甲基化酶2（DRM 2）和RNA聚合酶V（Pol V），这是RNA指导的DNA甲基化的两个主要作用者，但不依赖于Pol IV。我们的传感器以高时间分辨率在单细胞水平上提供了对全球DNA甲基化动态的洞察，并提供了一个强大的工具来跟踪CG和非CG甲基化，无论是在发育过程中还是在所有具有甲基化DNA的生物体中对环境线索的反应，正如我们在小鼠胚胎干细胞中所展示的那样。
In this Resource/Methodology, Ingouff et al. developed a new method to investigate DNA methylation reprogramming in plants. Using Arabidopsis, the authors developed sensors that reported CG and non-CG methylation with single-cell resolution. Cytosine methylation is a key epigenetic mark in many organisms, important for both transcriptional control and genome integrity. While relatively stable during somatic growth, DNA methylation is reprogrammed genome-wide during mammalian reproduction. Reprogramming is essential for zygotic totipotency and to prevent transgenerational inheritance of epimutations. However, the extent of DNA methylation reprogramming in plants remains unclear. Here, we developed sensors reporting with single-cell resolution CG and non-CG methylation in Arabidopsis. Live imaging during reproduction revealed distinct and sex-specific dynamics for both contexts. We found that CHH methylation in the egg cell depends on DOMAINS REARRANGED METHYLASE 2 (DRM2) and RNA polymerase V (Pol V), two main actors of RNA-directed DNA methylation, but does not depend on Pol IV. Our sensors provide insight into global DNA methylation dynamics at the single-cell level with high temporal resolution and offer a powerful tool to track CG and non-CG methylation both during development and in response to environmental cues in all organisms with methylated DNA, as we illustrate in mouse embryonic stem cells.