Institute of Molecular Genetics of the ASCR, v. v. i., Vídeňská 1083, 142 20 Prague 4, Czech Republic.
Max Planck Institute of Molecular Cell Biology and Genetics, 01307 Dresden, Germany.
Sci Rep. 2016 Jul 21;6:29961. doi: 10.1038/srep29961.
In search for the function of local chromatin environment on pre-mRNA processing we established a new tool, which allows for the modification of chromatin using a targeted approach. Using Transcription Activator-Like Effector domains fused to histone modifying enzymes (TALE-HME), we show locally restricted alteration of histone methylation modulates the splicing of target exons. We provide evidence that a local increase in H3K9 di- and trimethylation promotes inclusion of the target alternative exon, while demethylation by JMJD2D leads to exon skipping. We further demonstrate that H3K9me3 is localized on internal exons genome-wide suggesting a general role in splicing. Consistently, targeting of the H3K9 demethylase to a weak constitutive exon reduced co-transcriptional splicing. Together our data show H3K9 methylation within the gene body is a factor influencing recognition of both constitutive and alternative exons.
为了研究局部染色质环境对前体 mRNA 处理的功能，我们建立了一种新的工具，该工具允许通过靶向方法修饰染色质。我们使用融合了组蛋白修饰酶的转录激活因子样效应结构域（TALE-HME），证明了局部限制的组蛋白甲基化修饰可调节靶外显子的剪接。我们提供的证据表明，H3K9 二甲基化和三甲基化的局部增加促进了靶标选择性外显子的包含，而 JMJD2D 的去甲基化导致外显子跳过。我们进一步表明，H3K9me3 在内含子上在全基因组范围内定位，这表明其在剪接中具有普遍作用。一致地，将 H3K9 去甲基酶靶向弱组成性外显子可减少共转录剪接。总之，我们的数据表明，基因体中的 H3K9 甲基化是影响识别组成性和选择性外显子的因素之一。
