Functional Analysis of the Cathepsin D Gene Response to SGIV Infection in the Orange-Spotted Grouper, Epinephelus coioides
文献类型： 外文期刊
作者： Wang, Yuexuan 1 ; Han, Honglin 1 ; Zhu, Kecheng 2 ; Xu, Suifeng 1 ; Han, Chengzong 1 ; Jiang, Yunxiang 1 ; Wei, Shina 1 ; Qin, Qiwei 1 ;
作者机构： 1.South China Agr Univ, Coll Marine Sci, Lab Lingnan Modern Agr, Guangzhou 510642, Peoples R China
2.Chinese Acad Fishery Sci, South China Sea Fisheries Res Inst, Key Lab South China Sea Fishery Resources Exploit, Minist Agr & Rural Affairs, Guangzhou 510300, Peoples R China
3.Southern Marine Sci & Engn Guangdong Lab Zhuhai, Zhuhai 528478, Peoples R China
4.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Biol & Biotechnol, Qingdao 266000, Peoples R China
关键词： Singapore grouper iridovirus (SGIV); grouper; cathepsin D; apoptosis
期刊名称：VIRUSES-BASEL （ 影响因子：5.818； 五年影响因子：5.811 ）
ISSN：
年卷期： 2022 年 14 卷 8 期
页码：
收录情况： SCI
摘要： (1) Background: Lysosomal aspartic protease Cathepsin D (CD) is a key regulator and signaling molecule in various biological processes including activation and degradation of intracellular proteins, the antigen process and programmed cell death. However, the function of fish CD in virus infection remains largely unknown. (2) Methods: The functions of the CD gene response to SGIV infection was determined with light microscopy, reverse transcription quantitative PCR, Western blot and flow cytometry. (3) Results: In this study, Ec-Cathepsin D (Ec-CD) was cloned and identified from the orange-spotted grouper, Epinephelus coioides. The open reading frame (ORF) of Ec-CD consisted of 1191 nucleotides encoding a 396 amino acid protein with a predicted molecular mass of 43.17 kDa. Ec-CD possessed typical CD structural features including an N-terminal signal peptide, a propeptide region and a mature domain including two glycosylation sites and two active sites, which were conserved in other CD sequences. Ec-CD was predominantly expressed in the spleen and kidneys of healthy groupers. A subcellular localization assay indicated that Ec-CD was mainly distributed in the cytoplasm. Ec-CD expression was suppressed by SGIV stimulation and Ec-CD-overexpressing inhibited SGIV replication, SGIV-induced apoptosis, caspase 3/8/9 activity and the activation of reporter gene p53 and activating protein-1 (AP-1) in vitro. Simultaneously, Ec-CD overexpression obviously restrained the activated mitogen-activated protein kinase (MAPK) pathways, including extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK). In addition, Ec-CD overexpression negatively regulated the transcription level of pro-inflammatory cytokines and activation of the NF-kappa B promotor. (4) Conclusions: Our findings revealed that the Ec-CD possibly served a function during SGIV infection.
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