Genomic location of PRMT6-dependent H3R2 methylation is decisive for the transcriptional outcome of associated genes [ChIP-seq]. Genomic location of PRMT6-dependent H3R2 methylation is decisive for the transcriptional outcome of associated genes [ChIP-seq]

Protein arginine methyltransferase 6 (PRMT6) is an epigenetic regulator of fundamental cellular processes, such as gene expression and DNA repair. Asymmetric dimethylation of histone H3 at arginine 2 (H3R2me2a) is the major histone modification catalyzed by PRMT6. To identify the genome-wide deposition and transcriptional impact of H3R2me2a, we established PRMT6 deletion in a human cell model of neural differentiation. These knockout cells show severe neural differentiation defects. ChIP-seq profiling reveals that H3R2me2a is present at promoter as well as non-promoter sites in a PRMT6-dependent manner. Loss of H3R2me2a causes enhanced H3K4me3 deposition and target gene transcription supporting a genome-wide repressive nature of H3R2me2a. Intriguingly, the non-promoter H3R2me2a peaks co-localize with active enhancer marks, such as H3K4me1 and H3K27ac. Overall design: Examining of PRMT6 dependent H3R2me2a deposition in CRISPR control and knockout PRMT6 with neighboring histone modification.

蛋白质精氨酸甲基转移酶6（Protein arginine methyltransferase 6, PRMT6）是一类调控基因表达、DNA修复等基本细胞过程的表观遗传调控因子。由PRMT6催化的核心组蛋白修饰为组蛋白H3精氨酸2位点的不对称二甲基化（H3R2me2a）。为解析H3R2me2a的全基因组分布模式及其转录调控效应，本研究在神经分化的人类细胞模型中构建了PRMT6敲除体系。该敲除细胞系表现出严重的神经分化缺陷。染色质免疫共沉淀测序（ChIP-seq）谱分析显示，H3R2me2a以PRMT6依赖的方式分布于启动子及非启动子区域。H3R2me2a的缺失会导致H3K4me3修饰水平升高及靶基因转录增强，这证实了H3R2me2a在全基因组范围内的抑制性功能。值得注意的是，非启动子区域的H3R2me2a峰与H3K4me1、H3K27ac等活性增强子标记物存在共定位现象。实验整体设计：探究CRISPR对照与PRMT6敲除细胞中PRMT6依赖的H3R2me2a分布，及其伴随的邻近组蛋白修饰变化。