De novo induction of a DNA-histone H3K9 methylation loop on synthetic human repetitive DNA in cultured tobacco cells
De novo induction of a DNA-histone H3K9 methylation loop on synthetic human repetitive DNA in cultured tobacco cells
复制标题
DOI：
10.1111/tpj.16164
复制
发表时间：
2023-03-17
期刊：
影响因子：
7.2
通讯作者：
Masumoto，Hiroshi
中科院分区：
文献类型：
--
作者：
Otake，Koichiro;Kugou，Kazuto;Masumoto，Hiroshi
植物的遗传修饰是基础研究和应用研究的重要工具。转基因表达通常在独立品系或其后代中变化，并且与插入位点的染色质结构相关。基于理解如何操纵插入基因盒的表观遗传状态的策略将有助于确保转基因表达。在这里，我们报告了一种染色质操作的策略，通过人工拴系表观遗传效应子到植物亮黄-2（BY-2）培养细胞中的合成人着丝粒重复DNA（α DNA）平台。通过拴系DNA甲基转移酶（烟草DRM 1），我们在α DNA平台上有效地诱导了DNA甲基化和组蛋白甲基化（H3 K9 me 2）。据报道缺乏组蛋白甲基转移酶活性的拟南芥SUVH 9的拴系也诱导了BY-2细胞中α DNA上类似的表观遗传状态，推测是通过激活RNA依赖性DNA甲基化（RdDM）途径。我们的研究结果强调DNA和组蛋白甲基化机制之间的相互作用是植物细胞固有的。我们还发现，一旦表观遗传修饰状态被DRM 1或SUV H9的拴系诱导，即使当效应子的直接拴系被抑制时，该修饰也被维持。我们的系统能够分析更多样化的表观遗传效应子，并将有助于阐明植物细胞的染色质组装机制。
Genetic modifications in plants are crucial tools for fundamental and applied research. Transgene expression usually varies among independent lines or their progeny and is associated with the chromatin structure of the insertion site. Strategies based on understanding how to manipulate the epigenetic state of the inserted gene cassette would help to ensure transgene expression. Here, we report a strategy for chromatin manipulation by the artificial tethering of epigenetic effectors to a synthetic human centromeric repetitive DNA (alphoid DNA) platform in plant Bright‐Yellow‐2 (BY‐2) culture cells. By tethering DNA‐methyltransferase (Nicotiana tabacumDRM1), we effectively induced DNA methylation and histone methylation (H3K9me2) on the alphoid DNA platform. Tethering of the Arabidopsis SUVH9, which has been reported to lack histone methyltransferase activity, also induced a similar epigenetic state on the alphoid DNA in BY‐2 cells, presumably by activating the RNA‐dependent DNA methylation (RdDM) pathway. Our results emphasize that the interplay between DNA and histone methylation mechanisms is intrinsic to plant cells. We also found that once epigenetic modification states were induced by the tethering of either DRM1 or SUVH9, the modification was maintained even when the direct tethering of the effector was inhibited. Our system enables the analysis of more diverse epigenetic effectors and will help to elucidate the chromatin assembly mechanisms of plant cells.