Virus-induced p38 MAPK activation facilitates viral infection.
Virus-induced p38 MAPK activation facilitates viral infection.
复制标题
病毒诱导的 p38 MAPK 激活促进病毒感染
作者：
Cheng Y;Sun F;Wang L;Gao M;Xie Y;Sun Y;Liu H;Yuan Y;Yi W;Huang Z;Yan H;Peng K;Wu Y;Cao Z
理由：已知许多病毒感染会激活 p38 丝裂原激活蛋白激酶 (MAPK) 信号通路。然而，p38 激活在病毒感染中的作用及其潜在机制仍不清楚。本研究调查了病毒劫持的 p38 MAPK 激活在病毒感染中的作用。方法：通过免疫组织化学和蛋白质印迹研究患者组织和原代人肝细胞（PHH）中丙型肝炎病毒（HCV）感染与p38激活的相关性。使用免疫共沉淀、GST Pulldown 和共聚焦显微镜研究 p38α 和 HCV 核心蛋白的相互作用。使用体外激酶测定和质谱分析HCV核心蛋白的磷酸化。使用噬菌斑测定、定量实时 PCR (qRT-PCR)、蛋白质印迹、siRNA 和 CRISPR/Cas9 来确定 p38 激活对病毒复制的影响。结果：HCV 感染与临床样本中的 p38 激活相关。 HCV 感染通过触发 p38α 和 TGF-β 激活激酶 1 (MAP3K7) 结合蛋白 1 (TAB1) 的相互作用来增加 p38 磷酸化。 TAB1介导的p38α激活促进HCV复制，而SB203580对p38α激活的药物抑制抑制了病毒组装步骤中的HCV感染。激活的p38α与HCV核心蛋白的N端区域相互作用，随后磷酸化HCV核心蛋白，从而促进HCV核心蛋白寡聚化，这是病毒组装的重要步骤。正如预期的那样，SB203580 或 HCV 核心蛋白 N 末端肽（CN 肽）破坏了 p38α-HCV 核心蛋白相互作用，有效破坏了 HCV 组装，并阻碍了培养细胞和原代人肝细胞中的正常 HCV 复制。同样，严重发热伴血小板减少综合征病毒 (SFTSV)、单纯疱疹病毒 1 型 (HSV-1) 或严重急性呼吸综合征冠状病毒 2 (SARS-CoV-2) 感染也会激活 p38 MAPK。最重要的是，SB203580 对 p38 激活的药理学阻断有效抑制了 SFTSV、HSV-1 和 SARS-CoV-2。结论：我们的研究表明，病毒劫持的 p38 激活是病毒复制的关键事件，而药物阻断 p38 激活是一种抗病毒策略。
Rationale: Many viral infections are known to activate the p38 mitogen-activated protein kinase (MAPK) signaling pathway. However, the role of p38 activation in viral infection and the underlying mechanism remain unclear. The role of virus-hijacked p38 MAPK activation in viral infection was investigated in this study. Methods: The correlation of hepatitis C virus (HCV) infection and p38 activation was studied in patient tissues and primary human hepatocytes (PHHs) by immunohistochemistry and western blotting. Coimmunoprecipitation, GST pulldown and confocal microscopy were used to investigate the interaction of p38α and the HCV core protein. In vitro kinase assays and mass spectrometry were used to analyze the phosphorylation of the HCV core protein. Plaque assays, quantitative real time PCR (qRT-PCR), western blotting, siRNA and CRISPR/Cas9 were used to determine the effect of p38 activation on viral replication. Results: HCV infection was associated with p38 activation in clinical samples. HCV infection increased p38 phosphorylation by triggering the interaction of p38α and TGF-β activated kinase 1 (MAP3K7) binding protein 1 (TAB1). TAB1-mediated p38α activation facilitated HCV replication, and pharmaceutical inhibition of p38α activation by SB203580 suppressed HCV infection at the viral assembly step. Activated p38α interacted with the N-terminal region of the HCV core protein and subsequently phosphorylated the HCV core protein, which promoted HCV core protein oligomerization, an essential step for viral assembly. As expected, SB203580 or the HCV core protein N-terminal peptide (CN-peptide) disrupted the p38α-HCV core protein interaction, efficiently impaired HCV assembly and impeded normal HCV replication in both cultured cells and primary human hepatocytes. Similarly, severe fever with thrombocytopenia syndrome virus (SFTSV), herpes simplex virus type 1 (HSV-1) or severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection also activated p38 MAPK. Most importantly, pharmacological blockage of p38 activation by SB203580 effectively inhibited SFTSV, HSV-1 and SARS-CoV-2. Conclusion: Our study shows that virus-hijacked p38 activation is a key event for viral replication and that pharmacological blockage of p38 activation is an antiviral strategy.