Cloning of common carp SOCS-3 gene and its expression during embryogenesis, GH-transgene and viral infection.
As a member of a newly discovered protein family, the suppressor of cytokine signalling 3 (SOCS-3) has been shown to regulate the responses of many immune cytokines in a negative auto-regulatory manner. The full-length cDNA of common carp SOCS-3 was 1603 bp and contained a 630 bp open reading frame (ORF) coding for a protein of 209 amino acids. Carp SOCS-3 molecule was well conserved especially in the SRC homology 2 (SH2) and the SOCS box. The kinase inhibitory region (KIR) and ESS domains, upstream of the SH2 domain, were conserved in carp SOCS-3, except for a specific insertion (PHRYK) in the KIR domain at the N-terminal region. Three conserved cysteine (Cys-102, 124 and 193) residues, and one additional cysteine (Cys-168) residue, were also found in carp SOCS-3. The 2015 bp genomic DNA of carp SOCS-3 contained two exons and one intron. Phylogenetic analysis showed that carp SOCS-3 sequence grouped with other known fish SOCS-3 sequences with zebrafish SOCS-3 as the closest neighbour. RT-PCR analysis showed that carp SOCS-3 was initially expressed at 4 h pf (post-fertilization) and gradually increased up to 4 w pf during embryogenesis. By RT-qPCR analysis, carp SOCS-3 gene was predominantly detected in gill, head kidney, thymus and skin, followed by spleen and peripheral blood, lower expression level was detected in kidney, intestine, liver and muscle; the SOCS-3 transcript was significantly increased in thymus, head kidney, spleen and intestine of GH (growth hormone)-transgenic carp; after SVCV (spring viraemia of carp virus) infection, the carp SOCS-3 transcript was significantly up-regulated in gill, intestine, thymus, spleen, head kidney and kidney tissues in a time-dependent manner. These results suggest that teleost SOCS-3 may play an active role in the modulation of viral-induced innate immune response and in preventing the overaction of some cytokines with viral stimulation.
Xiao
Zhan-Gang
ZG
Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China.
Liu
Hong
H
Fu
Jian-Ping
JP
Hu
Wei
W
Wang
Ya-Ping
YP
Guo
Qiong-Lin
QL
D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D002347:Carps / Q000196:embryology / Q000235:genetics* / Q000276:immunology* / Q000821:virology; D003001:Cloning, Molecular; D047108:Embryonic Development / Q000276:immunology; D005393:Fish Diseases / Q000276:immunology*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D013006:Growth Hormone / Q000235:genetics; D008969:Molecular Sequence Data; D010802:Phylogeny; D018353:Rhabdoviridae Infections / Q000276:immunology / Q000662:veterinary*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D050826:Suppressor of Cytokine Signaling Proteins / Q000235:genetics* / Q000276:immunology*; D018116:Vesiculovirus / Q000276:immunology
D050826:Suppressor of Cytokine Signaling Proteins; D013006:Growth Hormone