Functional Analysis of the Cathepsin D Gene Response to SGIV Infection in the Orange-Spotted Grouper, <i>Epinephelus coioides</i>
(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 re...
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MDPI AG
2022-07-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/14/8/1680 |
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| author | Yuexuan Wang Honglin Han Kecheng Zhu Suifeng Xu Chengzong Han Yunxiang Jiang Shina Wei Qiwei Qin |
| author_facet | Yuexuan Wang Honglin Han Kecheng Zhu Suifeng Xu Chengzong Han Yunxiang Jiang Shina Wei Qiwei Qin |
| author_sort | Yuexuan Wang |
| collection | DOAJ |
| description | (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, <i>Epinephelus coioides</i>. 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-κB promotor. (4) Conclusions: Our findings revealed that the Ec-CD possibly served a function during SGIV infection. |
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| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-09T12:23:19Z |
| publishDate | 2022-07-01 |
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| series | Viruses |
| spelling | doaj.art-f8546a7015e84d9eb4d8b264584029942023-11-30T22:38:47ZengMDPI AGViruses1999-49152022-07-01148168010.3390/v14081680Functional Analysis of the Cathepsin D Gene Response to SGIV Infection in the Orange-Spotted Grouper, <i>Epinephelus coioides</i>Yuexuan Wang0Honglin Han1Kecheng Zhu2Suifeng Xu3Chengzong Han4Yunxiang Jiang5Shina Wei6Qiwei Qin7Laboratory for Lingnan Modern Agriculture, College of Marine Sciences, South China Agricultural University, Guangzhou 510642, ChinaLaboratory for Lingnan Modern Agriculture, College of Marine Sciences, South China Agricultural University, Guangzhou 510642, ChinaKey Laboratory of South China Sea Fishery Resources Exploitation and Utilization, Ministry of Agriculture and Rural Affairs, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510300, ChinaLaboratory for Lingnan Modern Agriculture, College of Marine Sciences, South China Agricultural University, Guangzhou 510642, ChinaLaboratory for Lingnan Modern Agriculture, College of Marine Sciences, South China Agricultural University, Guangzhou 510642, ChinaLaboratory for Lingnan Modern Agriculture, College of Marine Sciences, South China Agricultural University, Guangzhou 510642, ChinaLaboratory for Lingnan Modern Agriculture, College of Marine Sciences, South China Agricultural University, Guangzhou 510642, ChinaLaboratory for Lingnan Modern Agriculture, College of Marine Sciences, South China Agricultural University, Guangzhou 510642, China(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, <i>Epinephelus coioides</i>. 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-κB promotor. (4) Conclusions: Our findings revealed that the Ec-CD possibly served a function during SGIV infection.https://www.mdpi.com/1999-4915/14/8/1680Singapore grouper iridovirus (SGIV)groupercathepsin Dapoptosis |
| spellingShingle | Yuexuan Wang Honglin Han Kecheng Zhu Suifeng Xu Chengzong Han Yunxiang Jiang Shina Wei Qiwei Qin Functional Analysis of the Cathepsin D Gene Response to SGIV Infection in the Orange-Spotted Grouper, <i>Epinephelus coioides</i> Viruses Singapore grouper iridovirus (SGIV) grouper cathepsin D apoptosis |
| title | Functional Analysis of the Cathepsin D Gene Response to SGIV Infection in the Orange-Spotted Grouper, <i>Epinephelus coioides</i> |
| title_full | Functional Analysis of the Cathepsin D Gene Response to SGIV Infection in the Orange-Spotted Grouper, <i>Epinephelus coioides</i> |
| title_fullStr | Functional Analysis of the Cathepsin D Gene Response to SGIV Infection in the Orange-Spotted Grouper, <i>Epinephelus coioides</i> |
| title_full_unstemmed | Functional Analysis of the Cathepsin D Gene Response to SGIV Infection in the Orange-Spotted Grouper, <i>Epinephelus coioides</i> |
| title_short | Functional Analysis of the Cathepsin D Gene Response to SGIV Infection in the Orange-Spotted Grouper, <i>Epinephelus coioides</i> |
| title_sort | functional analysis of the cathepsin d gene response to sgiv infection in the orange spotted grouper i epinephelus coioides i |
| topic | Singapore grouper iridovirus (SGIV) grouper cathepsin D apoptosis |
| url | https://www.mdpi.com/1999-4915/14/8/1680 |
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