State Key Laboratory of Chemical Oncogenomics, Guangdong Provincial Key Laboratory of Chemical Genomics, Laboratory of Structural Biology and Drug Discovery, Laboratory of Ubiquitination and Targeted Therapy, School of Chemical Biology and Biotechnology, Peking University Shenzhen Graduate School, Shenzhen 518055, PR China.
National Clinical Research Center for Infectious Diseases, Shenzhen Third People's Hospital, Southern University of Science and Technology, Shenzhen 518112, PR China.
Sci Signal. 2023 May 2;16(783):eadd0082. doi: 10.1126/scisignal.add0082.
The SARS-CoV-2 papain-like protease (PLpro), which has deubiquitinating activity, suppresses the type I interferon (IFN-I) antiviral response. We investigated the mechanism by which PLpro antagonizes cellular antiviral responses. In HEK392T cells, PLpro removed K63-linked polyubiquitin chains from Lys of the stimulator of interferon genes (STING). PLpro-mediated deubiquitination of STING disrupted the STING-IKKε-IRF3 complex that induces the production of IFN-β and IFN-stimulated cytokines and chemokines. In human airway cells infected with SARS-CoV-2, the combined treatment with the STING agonist diABZi and the PLpro inhibitor GRL0617 resulted in the synergistic inhibition of SARS-CoV-2 replication and increased IFN-I responses. The PLpros of seven human coronaviruses (SARS-CoV-2, SARS-CoV, MERS-CoV, HCoV-229E, HCoV-HKU1, HCoV-OC43, and HCoV-NL63) and four SARS-CoV-2 variants of concern (α, β, γ, and δ) all bound to STING and suppressed STING-stimulated IFN-I responses in HEK293T cells. These findings reveal how SARS-CoV-2 PLpro inhibits IFN-I signaling through STING deubiquitination and a general mechanism used by seven human coronaviral PLpros to dysregulate STING and to facilitate viral innate immune evasion. We also identified simultaneous pharmacological STING activation and PLpro inhibition as a potentially effective strategy for antiviral therapy against SARS-CoV-2.
严重急性呼吸综合征冠状病毒 2 型（SARS-CoV-2）的木瓜样蛋白酶（PLpro）具有去泛素化活性，可抑制 I 型干扰素（IFN-I）抗病毒反应。我们研究了 PLpro 拮抗细胞抗病毒反应的机制。在 HEK392T 细胞中，PLpro 从干扰素基因刺激因子（STING）的赖氨酸上除去 K63 连接的多泛素链。PLpro 介导的 STING 去泛素化破坏了 STING-IKKε-IRF3 复合物，该复合物诱导 IFN-β 和 IFN 刺激的细胞因子和趋化因子的产生。在感染 SARS-CoV-2 的人呼吸道细胞中，STING 激动剂 diABZi 和 PLpro 抑制剂 GRL0617 的联合治疗导致 SARS-CoV-2 复制的协同抑制和 IFN-I 反应的增加。七种人类冠状病毒（SARS-CoV-2、SARS-CoV、MERS-CoV、HCoV-229E、HCoV-HKU1、HCoV-OC43 和 HCoV-NL63）和四种 SARS-CoV-2 关注变体（α、β、γ和δ）的 PLpros 都与 STING 结合，并抑制 HEK293T 细胞中 STING 刺激的 IFN-I 反应。这些发现揭示了 SARS-CoV-2 PLpro 如何通过 STING 去泛素化抑制 IFN-I 信号传导，以及七种人类冠状病毒 PLpros 用于失调 STING 和促进病毒先天免疫逃避的一般机制。我们还确定了同时进行药理学 STING 激活和 PLpro 抑制作为针对 SARS-CoV-2 的抗病毒治疗的潜在有效策略。
