Symmetric arginine dimethylation is selectively required for mRNA splicing and the initiation of type I and type III interferon signaling

The goal of this study was to assess the extent of symmetric arginine dimethylation-dependent splicing and its associated signaling outcomes by using human T lymphocyte activation as a model system with a high rate of de novo transcription. Naïve human T lymphocytes were activated and treated with a selective inhibitor of PRMT5, PF-06829927, and investigated for changes in gene expression, alternative splicing, and chromatin accessibility. Overall design: Human naïve CD3+ (ATAC), CD4+ or CD8+ (expression and splicing) T lymphocytes were activated for 72 (expression and splicing) or 96 (ATAC) hours with platebound anti-CD3, anti-CD28, IL-2, and either DMSO or PF-06829927

本研究旨在以新生转录速率极高的人类T淋巴细胞活化作为模型系统，评估精氨酸对称二甲基化依赖型可变剪接的发生范围及其相关信号传导结局。研究人员对初始态人类T淋巴细胞进行活化处理，并施加蛋白精氨酸甲基转移酶5（PRMT5）选择性抑制剂PF-06829927，随后探究其基因表达、可变剪接及染色质开放状态的变化。实验总体设计如下：将人类初始态CD3+ T淋巴细胞（用于转座酶可及性染色质测序（ATAC））、CD4+或CD8+ T淋巴细胞（用于基因表达与可变剪接分析），分别经贴壁包被的抗CD3、抗CD28抗体及IL-2刺激活化72小时（用于基因表达与剪接分析）或96小时（用于ATAC测序），同时以二甲基亚砜（DMSO）或PF-06829927进行处理。