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./nMethods: 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./nResults: 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./nConclusion: 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.

中文翻译：

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病毒诱导的p38 MAPK活化促进病毒感染

基本原理：已知许多病毒感染会激活p38丝裂原激活的蛋白激酶（MAPK）信号传导途径。但是，p38激活在病毒感染中的作用及其潜在机制尚不清楚。这项研究研究了病毒劫持的p38 MAPK激活在病毒感染中的作用。/n方法：通过免疫组织化学和免疫组织化学方法研究了患者组织和原代人肝细胞（PHHs）中丙型肝炎病毒（HCV）感染与p38激活的相关性。蛋白质印迹。使用共免疫沉淀，GST下拉和共聚焦显微镜研究p38α和HCV核心蛋白的相互作用。体外激酶测定和质谱用于分析HCV核心蛋白的磷酸化。噬菌斑测定，定量实时PCR（QRT-PCR），western印迹，siRNA和CRISPR / Cas9用于测定p38活化对病毒replication./n的效果结果：HCV感染与临床样本中的p38激活有关。HCV感染通过触发p38α和TGF-β活化激酶1（MAP3K7）结合蛋白1（TAB1）的相互作用而增加p38磷酸化。TAB1介导的p38α激活促进了HCV复制，而SB203580对p38α激活的药物抑制作用抑制了病毒装配步骤中的HCV感染。活化的p38α与HCV核心蛋白的N末端区域相互作用，随后使HCV核心蛋白磷酸化，从而促进HCV核心蛋白寡聚化，这是病毒装配的重要步骤。如预期的那样，SB203580或HCV核心蛋白N末端肽（CN肽）破坏了p38α-HCV核心蛋白的相互作用，有效地破坏了HCV装配并阻碍了HCV在培养细胞和原代人肝细胞中的复制。同样，患有血小板减少症综合征病毒（SFTSV），1型单纯疱疹病毒（HSV-1）或严重急性呼吸综合征冠状病毒2（SARS-CoV-2）感染的严重发烧也激活了p38 MAPK。最重要的是，SB203580对p38激活的药理阻断作用可有效抑制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p38激活的药理学阻断作用是一种抗病毒策略。