Department of Immunology, School of Basic Medical Sciences, Beijing Key Laboratory of Tumor Systems Biology, Institute of Systems Biomedicine, Peking University Health Science Center, Beijing, China.
Beijing Institutes of Life Science, Chinese Academy of Sciences, Beijing, China.
PLoS Pathog. 2022 Jun 3;18(6):e1010599. doi: 10.1371/journal.ppat.1010599. eCollection 2022 Jun.
Regulation of chromatin structure and accessibility determines the transcription activities of genes, which endows the host with function-specific patterns of gene expression. Upon viral infection, the innate immune responses provide the first line of defense, allowing rapid production of variegated antiviral cytokines. Knowledge on how chromatin accessibility is regulated during host defense against viral infection remains limited. Our previous work found that the nuclear matrix protein SAFA surveilled viral RNA and regulated antiviral immune genes expression. However, how SAFA regulates the specific induction of antiviral immune genes remains unknown. Here, through integration of RNA-seq, ATAC-seq and ChIP-seq assays, we found that the depletion of SAFA specifically decreased the chromatin accessibility, activation and expression of virus induced genes. And mutation assays suggested that the RNA-binding ability of SAFA was essential for its function in regulating antiviral chromatin accessibility. RIP-seq results showed that SAFA exclusively bound with antiviral related RNAs following viral infection. Further, we combined the CRISPR-Cas13d mediated RNA knockdown system with ATAC-qPCR, and demonstrated that the binding between SAFA and according antiviral RNAs specifically mediated the openness of the corresponding chromatin and following robust transcription of antiviral genes. Moreover, knockdown of these associated RNAs dampened the accessibility of related genes in an extranuclear signaling pathway dependent manner. Interestingly, VSV infection cleaved SAFA protein at the C-terminus which deprived its RNA binding ability for immune evasion. Thus, our results demonstrated that SAFA and the interacting RNA products collaborated and remodeled chromatin accessibility to facilitate antiviral innate immune responses.
染色质结构和可及性的调节决定了基因的转录活性,这使宿主具有特定功能的基因表达模式。在病毒感染时,先天免疫反应提供了第一道防线,允许快速产生多样化的抗病毒细胞因子。关于宿主防御病毒感染时染色质可及性如何调节的知识仍然有限。我们之前的工作发现,核基质蛋白 SAFA 监测病毒 RNA 并调节抗病毒免疫基因的表达。然而,SAFA 如何调节抗病毒免疫基因的特异性诱导仍然未知。在这里,通过整合 RNA-seq、ATAC-seq 和 ChIP-seq 分析,我们发现 SAFA 的耗竭特异性降低了病毒诱导基因的染色质可及性、激活和表达。突变分析表明,SAFA 的 RNA 结合能力对于其调节抗病毒染色质可及性的功能至关重要。RIP-seq 结果表明,SAFA 在病毒感染后仅与抗病毒相关的 RNA 结合。此外,我们将 CRISPR-Cas13d 介导的 RNA 敲低系统与 ATAC-qPCR 相结合,证明了 SAFA 与相应抗病毒 RNA 之间的结合特异性介导了相应染色质的开放性,随后抗病毒基因的转录增强。此外,这些相关 RNA 的敲低以依赖于核外信号通路的方式抑制了相关基因的可及性。有趣的是,VSV 感染在 C 端切割 SAFA 蛋白,使其丧失了免疫逃避的 RNA 结合能力。因此,我们的结果表明,SAFA 和相互作用的 RNA 产物协同作用并重塑染色质可及性,以促进抗病毒先天免疫反应。