PUBLICATION
Characterization of c-Jun from orange-spotted grouper, Epinephelus coioides involved in SGIV infection
- Authors
- Wei, S., Huang, Y., Huang, X., Qin, Q.
- ID
- ZDB-PUB-150104-4
- Date
- 2015
- Source
- Fish & shellfish immunology 43(1): 230-40 (Journal)
- Registered Authors
- Keywords
- DN-Ec-c-Jun, Epinephelus coioides, Singapore grouper iridovirus (SGIV), c-Jun, virus replication
- MeSH Terms
-
- Amino Acid Sequence
- Animals
- Base Sequence
- Bass*
- DNA Virus Infections/genetics
- DNA Virus Infections/veterinary*
- DNA Virus Infections/virology
- Fish Diseases/genetics*
- Fish Diseases/virology
- Fish Proteins/chemistry
- Fish Proteins/genetics*
- Fish Proteins/metabolism
- Iridovirus/physiology*
- Molecular Sequence Data
- Phylogeny
- Proto-Oncogene Proteins c-jun/chemistry
- Proto-Oncogene Proteins c-jun/genetics*
- Proto-Oncogene Proteins c-jun/metabolism
- PubMed
- 25555808 Full text @ Fish Shellfish Immunol.
Citation
Wei, S., Huang, Y., Huang, X., Qin, Q. (2015) Characterization of c-Jun from orange-spotted grouper, Epinephelus coioides involved in SGIV infection. Fish & shellfish immunology. 43(1):230-40.
Abstract
The 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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping