The substrate selectivity of papain-like proteases from human-infecting coronaviruses correlates with innate immune suppression
The substrate selectivity of papain-like proteases from human-infecting coronaviruses correlates with innate immune suppression
复制标题
DOI：
10.1126/scisignal.ade1985
复制
发表时间：
2023-05
影响因子：
7.3
通讯作者：
Yuxian Xiong;Bing Huang;Ying Yang;Xinming Fu;Ziyang Fu;Huidong Xu;Ming Liu;D. Cao;Manman Zhang;Haibin Yang;Xiaogang Niu;Cong Yu;Hao Huang
中科院分区：
文献类型：
--
作者：
Yuxian Xiong;Bing Huang;Ying Yang;Xinming Fu;Ziyang Fu;Huidong Xu;Ming Liu;D. Cao;Manman Zhang;Haibin Yang;Xiaogang Niu;Cong Yu;Hao Huang
可感染人类的冠状病毒可引起普通感冒（HCoV-NL63、HCoV-229E、HCoV-HKU1 和 HCoV-OC43）或严重呼吸道症状（SARS-CoV-2、SARS-CoV 和 MERS-CoV）。 SARS-CoV、SARS-CoV-2、MERS-CoV 和 HCoV-NL63 的木瓜蛋白酶 (PLP) 在病毒先天免疫逃避中发挥作用，并具有去泛素化 (DUB) 和去糖基化活性。我们鉴定了 HCoV-229E、HCoV-HKU1 和 HCoV-OC43 的 PLP，并发现它们的酶特性与其抑制先天免疫反应的能力相关。保守的非催化天冬氨酸残基对于 DUB 和去ISG化活性至关重要，但 PLP 对 Ub、K48 连接的 diUb 和干扰素刺激基因 15 (ISG15) 底物具有不同的泛素 (Ub) 链裂解选择性和结合亲和力。与 Ub 复合的 HKU1-PLP2 的晶体结构揭示了结合界面，这解释了该 PLP 和 Ub 之间异常高的结合亲和力。在细胞检测中，来自严重致病冠状病毒的PLP强烈抑制先天免疫IFN-I和NF-κB信号传导并刺激自噬，而来自轻度致病冠状病毒的PLP通常对免疫抑制和自噬诱导作用较弱。此外，来自关注的 SARS-CoV-2 变体的 PLP 显示出对先天免疫信号通路的抑制增强。总体而言，这些结果表明，这些 PLP 的 DUB 和去糖基化活性以及底物选择性对病毒先天免疫逃避有不同的贡献，并可能影响病毒的致病性。描述 蛋白酶对泛素或 ISG15 的偏好与冠状病毒的致病性相关。木瓜蛋白酶和冠状病毒致病性来自高致病性冠状病毒（包括 SARS-CoV-2）的木瓜蛋白酶 (PLP) 具有去泛素化和去糖基化活性，使其能够干扰先天免疫信号通路。 Xiong等人通过鉴定和表征弱病原性冠状病毒PLP的去泛素化和去ISG化活性。揭示了 PLP 的酶特性与致病性之间的关系。与来自高致病性冠状病毒的PLP相比，来自轻度致病性冠状病毒的PLP更喜欢泛素而不是ISG15。这种底物偏好的结构基础是由弱病原体 HCoV-HKU1 的 PLP2 晶体结构揭示的。尽管所有 PLP 都不同程度地抑制先天免疫信号通路，但来自高致病性病毒的 PLP 抑制作用更强烈，包括来自 SARS-CoV-2 变种的 PLP。这些差异可能导致冠状病毒之间免疫逃避和致病性的差异。 —AMV
Coronaviruses that can infect humans can cause either common colds (HCoV-NL63, HCoV-229E, HCoV-HKU1, and HCoV-OC43) or severe respiratory symptoms (SARS-CoV-2, SARS-CoV, and MERS-CoV). The papain-like proteases (PLPs) of SARS-CoV, SARS-CoV-2, MERS-CoV, and HCoV-NL63 function in viral innate immune evasion and have deubiquitinating (DUB) and deISGylating activities. We identified the PLPs of HCoV-229E, HCoV-HKU1, and HCoV-OC43 and found that their enzymatic properties correlated with their ability to suppress innate immune responses. A conserved noncatalytic aspartic acid residue was critical for both DUB and deISGylating activities, but the PLPs had differing ubiquitin (Ub) chain cleavage selectivities and binding affinities for Ub, K48-linked diUb, and interferon-stimulated gene 15 (ISG15) substrates. The crystal structure of HKU1-PLP2 in complex with Ub revealed binding interfaces that accounted for the unusually high binding affinity between this PLP and Ub. In cellular assays, the PLPs from the severe disease–causing coronaviruses strongly suppressed innate immune IFN-I and NF-κB signaling and stimulated autophagy, whereas the PLPs from the mild disease–causing coronaviruses generally showed weaker effects on immune suppression and autophagy induction. In addition, a PLP from a SARS-CoV-2 variant of concern showed increased suppression of innate immune signaling pathways. Overall, these results demonstrated that the DUB and deISGylating activities and substrate selectivities of these PLPs differentially contribute to viral innate immune evasion and may affect viral pathogenicity. Description The preference of a protease for ubiquitin or ISG15 is associated with coronavirus pathogenicity. Papain-like proteases and coronavirus pathogenicity The papain-like proteases (PLPs) from highly pathogenic coronaviruses, including SARS-CoV-2, have deubiquitylating and deISGylating activities that enable them to interfere with innate immune signaling pathways. By identifying and characterizing the deubiquitylation and deISGylation activities of PLPs from weakly pathogenic coronaviruses, Xiong et al. uncovered a relationship between the enzymatic properties of PLPs and pathogenicity. In contrast to those from the highly pathogenic coronaviruses, PLPs from the mildly pathogenic coronaviruses preferred ubiquitin over ISG15. The structural basis of this substrate preference was revealed by a crystal structure of PLP2 from the weak pathogen HCoV-HKU1. Although all PLPs inhibited innate immune signaling pathways to varying extents, those from the highly pathogenic viruses did so more strongly, including the PLP from a SARS-CoV-2 variant of concern. These differences likely contribute to the differences in immune evasion and pathogenicity among the coronaviruses. —AMV