Ubiquitin-specific protease 5 promotes EV-A71 replication by de-ubiquitinating MAVS and IRF3

Shumin Zhang; Yuan Fang; Shuai Ren; Xuhua Zhang; Chenggong Zheng; Zhipeng Qin; Wenqiang Wei; Huabin Zheng; Chuntian Li; Zekun Wang; Yujie Ren

doi:10.1016/j.virs.2025.12.002

December 2025

Citation: Shumin Zhang, Yuan Fang, Shuai Ren, Xuhua Zhang, Chenggong Zheng, Zhipeng Qin, Wenqiang Wei, Huabin Zheng, Chuntian Li, Zekun Wang, Yujie Ren. Ubiquitin-specific protease 5 promotes EV-A71 replication by de-ubiquitinating MAVS and IRF3 .VIROLOGICA SINICA, 2025, 40(6) : 910-920. http://dx.doi.org/10.1016/j.virs.2025.12.002

Ubiquitin-specific protease 5 promotes EV-A71 replication by de-ubiquitinating MAVS and IRF3

Shumin Zhang ^{a
,,} ,
Yuan Fang ^b,c ,
Shuai Ren ^a ,
Xuhua Zhang ^b,c ,
Chenggong Zheng ^a ,
Zhipeng Qin ^a ,
Wenqiang Wei ^e ,
Huabin Zheng ^a,d ,
Chuntian Li ^a,d ,
Zekun Wang ^{a,f
,,} ,
Yujie Ren ^{b,c
,,}

a. Joint National Laboratory for Antibody Drug Engineering, Henan University, Kaifeng 475004, China;
b. State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China;
c. University of Chinese Academy of Sciences, Beijing 100049, China;
d. The First Affiliated Hospital of Henan University, Henan University, Kaifeng 475004, China;
e. School of Basic Medical Sciences, Henan University, Kaifeng 475004, China;
f. State Key Laboratory of Antiviral Drugs, Henan University, Kaifeng 475004, China

Corresponding author: Shumin Zhang, shuminzhang@henu.edu.cn
Zekun Wang, watearth@foxmail.com
Yujie Ren, renyujie@wh.iov.cn
Received Date: 11 April 2025
Accepted Date: 02 December 2025
Available online: 04 December 2025

Abstract

Human enterovirus A71 (EV-A71) is a major causative agent of hand, foot and mouth disease (HFMD), which poses a significant public health threat, particularly among young children. Mitochondrial antiviral signaling protein (MAVS) and interferon regulatory factor 3 (IRF3) are vital proteins for the induction of type I interferons (IFN-I) and downstream interferon-stimulated genes (ISGs) during EV-A71 infection. While posttranslational modifications are known to critically influence viral infection processes, the mechanisms by which EV-A71 exploits host deubiquitinases (DUBs) for immune evasion remain poorly understood. In this study, we demonstrated that EV-A71 infection upregulated ubiquitin-specific protease 5 (USP5) expression. Knockdown of USP5 not only inhibited EV-A71 replication but also observably increased the production of IFN-I and ISGs. Furthermore, USP5 also regulated the replication of EV-D68 and CVA16 and the production of IFN-I and ISGs. Mechanistically, USP5 physically interacted with MAVS and IRF3 and reduced the K63-linked polyubiquitination of MAVS and IRF3. Conversely, USP5 knockdown increased the K63-linked polyubiquitination of MAVS and IRF3, thereby accelerating the phosphorylation of IRF3 and increasing IFN-I production during EV-A71 infection. Furthermore, pharmacological inhibition of USP5 with the small-molecule inhibitor PR-619 significantly potentiated the antiviral effects of IFN against EV-A71. Collectively, our findings reveal a previously unrecognized role of USP5 in facilitating EV-A71 immune evasion by dampening MAVS- and IRF3-mediated antiviral signaling. These insights provide a novel therapeutic avenue for combating EV-A71 infection through targeted modulation of the USP5-IRF3 axis.

Electronic Supplementary Material

10.1016j.virs.2025.12.002-ESM1.docx
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