Grouper USP12 exerts antiviral activity against nodavirus infection

Ya Zhang; Liwei Huang; Xiaolin Gao; Qiwei Qin; Xiaohong Huang; Youhua Huang

doi:10.1016/j.fsi.2022.01.011

Grouper USP12 exerts antiviral activity against nodavirus infection

Fish Shellfish Immunol. 2022 Feb:121:332-341. doi: 10.1016/j.fsi.2022.01.011. Epub 2022 Jan 13.

Authors

Ya Zhang¹, Liwei Huang¹, Xiaolin Gao¹, Qiwei Qin², Xiaohong Huang³, Youhua Huang⁴

Affiliations

¹ University Joint Laboratory of Guangdong Province, Hong Kong and Macao Region on Marine Bioresource Conservation and Exploitation, College of Marine Sciences, South China Agricultural University, Guangzhou, 510642, China.
² University Joint Laboratory of Guangdong Province, Hong Kong and Macao Region on Marine Bioresource Conservation and Exploitation, College of Marine Sciences, South China Agricultural University, Guangzhou, 510642, China; Guangdong Laboratory for Lingnan Modern Agriculture, Guangzhou, 510642, China; Southern Marine Science and Engineering Guangdong Laboratory, Zhuhai, 519082, China; Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266000, China.
³ University Joint Laboratory of Guangdong Province, Hong Kong and Macao Region on Marine Bioresource Conservation and Exploitation, College of Marine Sciences, South China Agricultural University, Guangzhou, 510642, China. Electronic address: huangxh@scau.edu.cn.
⁴ University Joint Laboratory of Guangdong Province, Hong Kong and Macao Region on Marine Bioresource Conservation and Exploitation, College of Marine Sciences, South China Agricultural University, Guangzhou, 510642, China. Electronic address: huangyh@scau.edu.cn.

PMID: 35032679
DOI: 10.1016/j.fsi.2022.01.011

Abstract

The ubiquitin-specific proteases (USPs) have attracted particular attention due to their multiple functions in different biological processes. USP12, a member of the USP family, has been demonstrated to exert critical roles in diverse cellular processes, including cell death, cancer and antiviral immunity. Here, we cloned a USP12 homolog from orange spotted grouper (Epinephelus coioides, E. coioides), and its roles in fish RNA virus replication were investigated. EcUSP12 contained a 1119-bp open reading frame (ORF) encoding a 372-amino acid polypeptide, which shared 100.00% and 91.32% identity with USP12 homolog of Etheostoma cragini and Homo sapiens, respectively. Sequence analysis indicated that EcUSP12 contained a conserved peptidase-C19G domain (aa 40-369). qPCR analysis showed that EcUSP12 transcript was most abundant in head kidney and spleen of grouper E. coioides. The expression of EcUSP12 was significantly upregulated in grouper spleen (GS) cells in response to red-spotted grouper nervous necrosis virus (RGNNV) infection. Subcellular localization analysis showed that EcUSP12 was evenly distributed throughout the cytoplasm, and mainly co-localized with endoplasmic reticulum (ER). Interestingly, during RGNNV infection, the endogenous distribution of EcUSP12 was obviously altered, and mostly overlapped with viral coat protein (CP). Co-Immunoprecipitation (Co-IP) assay indicated that EcUSP12 interacted with viral CP. In addition, overexpression of EcUSP12 significantly inhibited the replication of RGNNV in vitro, as evidenced by the decrease in viral gene transcription and protein synthesis during infection. Consistently, knockdown of EcUSP12 by small interfering RNA (siRNA) promoted the replication of RGNNV. Furthermore, EcUSP12 overexpression also increased the transcription level of inflammatory factors and interferon-related genes, including tumor necrosis factor α (TNF-α), interleukin (IL)-1β, IL-6, IL-8, interferon regulatory factor 3 (IRF3), and IRF7. Taken together, our results demonstrated that EcUSP12, as a positive regulator of IFN signaling, interacted with viral CP to inhibit virus infection.

Keywords: Coat protein; Epinephelus coioides; Immune responses; RGNNV; USP12.

MeSH terms

Amino Acid Sequence
Animals
Bass* / immunology
Bass* / virology
DNA Virus Infections* / immunology
DNA Virus Infections* / veterinary
Fish Diseases* / virology
Fish Proteins / immunology*
Immunity, Innate*
Nodaviridae
Phylogeny
Sequence Alignment
Ubiquitin-Specific Proteases / immunology*

Substances

Fish Proteins
Ubiquitin-Specific Proteases