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Characterization of c-Jun from orange-spotted grouper, Epinephelus coioides involved in SGIV infection
Wei, Shina1; Huang, Youhua1; Huang, Xiaohong1; Qin, Qiwei1;;
AbstractThe nuclear phosphoprotein c-Jun is a member of the AP1 family of transcription activating complex, can be induced by various extracellular stimuli such as virus infection. In this study, the c-Jun gene (Ec-c-Jun) was cloned from orange-spotted grouper, Epinephelus coioides. The full-length Ec-c-Jun cDNA is composed of 2046 bp and encodes a polypeptide of 328 amino acids with 81% identity of zebrafish. Amino acid alignment analysis indicated that Ec-c-Jun contained three conserved domains including a transactivation domain (TAD), a DNA-binding domain (DBD) and leucine zipper domain (LZD). RT-PCR results showed that Ec-c-Jun transcript was most abundant in spleen, kidney, heart and gill. The expression of Ec-c-Jun was up-regulated after challenged with Singapore grouper iridovirus (SGIV). To investigate the roles of Ec-c-Jun during SGIV infection, we constructed its dominant-negative mutant (DN-Ec-c-Jun) by deleting the major TAD that lacks amino acids 3-122. Fluorescence microscopy observation revealed that Ec-c-Jun and DN-Ec-c-Jun were expressed predominantly in the nucleus in transfected cells. Interestingly, the green fluorescence of Ec-c-Jun was congregated and co-localized with virus assembly sites at the late stage of SGIV infection. However, in DN-Ec-c-Jun transfected cells, no virus assembly sites were observed, and the distribution of fluorescence remained unchanged. Moreover, overexpression of DN-Ec-c-Jun in vitro delayed the occurrence of CPE induced by SGIV infection and inhibited the virus gene transcription. In addition, ectopic expression of DN-Ec-c-Jun was able to inhibit SGIV induced c-Jun/AP1 promoter activity in GS cells. Thus, we proposed that c-Jun transcription factor was essential for SGIV replication in vitro. Our results will contribute to understanding the crucial roles of JNK signaling pathway in fish virus infection. (C) 2014 Elsevier Ltd. All rights reserved.
KeywordEpinephelus Coioides C-jun Dn-ec-c-jun Singapore Grouper Iridovirus (Sgiv) Virus Replication
Subject AreaFisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
Document Type期刊论文
Affiliation1.[Wei, Shina
2.Huang, Youhua
3.Huang, Xiaohong
4.Qin, Qiwei] Chinese Acad Sci, South China Sea Inst Oceanol, Key Lab Trop Marine Bioresources & Ecol, Guangzhou 510301, Guangdong, Peoples R China
5.[Wei, Shina
6.Huang, Youhua
7.Huang, Xiaohong
8.Qin, Qiwei] Univ Chinese Acad Sci, Beijing, Peoples R China
Recommended Citation
GB/T 7714
Wei, Shina,Huang, Youhua,Huang, Xiaohong,et al. Characterization of c-Jun from orange-spotted grouper, Epinephelus coioides involved in SGIV infection[J]. FISH & SHELLFISH IMMUNOLOGY,2015,43(1):230-240.
APA Wei, Shina,Huang, Youhua,Huang, Xiaohong,Qin, Qiwei,,& of c-Jun from orange-spotted grouper, Epinephelus coioides involved in SGIV infection.FISH & SHELLFISH IMMUNOLOGY,43(1),230-240.
MLA Wei, Shina,et al."Characterization of c-Jun from orange-spotted grouper, Epinephelus coioides involved in SGIV infection".FISH & SHELLFISH IMMUNOLOGY 43.1(2015):230-240.
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