Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
5540270 | Fish & Shellfish Immunology | 2017 | 9 Pages |
Abstract
Interferon gamma (IFN-γ) is a major component in immunological signaling and plays a key role in resisting viral infection. In this study, we identified and characterized an IFN-γ gene (AsIFN-γ) in the marine fish black seabream (Acanthopagrus schlegelii). We cloned AsIFN-γ genomic sequence, which comprises four exons, three introns and an upstream promoter including several conserved regulatory elements. The complete cDNA of AsIFN-γ was 816 bp in length and encoded a putative 194 amino acids (aa) protein with a 22 aa signal peptide, six α-helices and one nuclear localization signal (NLS). Multiple alignment showed that AsIFN-γ protein shared 31-60% identity with IFN-γ of other fish but low identity with fish IFN-γrel and IFN-γ of other vertebrates. AsIFN-γ was constitutively expressed in all examined tissues with the highest expression level in immune organs, such as spleen, gill and kidney. In black seabream infected by red spotted nervous necrosis virus (RGNNV), the expression of AsIFN-γ was significantly up-regulated in most tissues, and RGNNV infection in vitro also induced significant up-regulation of AsIFN-γ, indicating that AsIFN-γ was involved in immune response to RGNNV infection. Overexpression of AsIFN-γ in cultured Acanthopagrus schlegelii brain (AsB) cells rapidly and transiently stimulated the expression of JAK-STAT signaling pathway related genes including STAT1, STAT2 and IRF9, as well as the downstream antiviral genes MX1 and ISG15. Furthermore, overexpression of AsIFN-γ was able to significantly inhibit RGNNV replication and virus production in AsB cells. In summary, we identified a conserved IFN-γ gene of black seabream, and demonstrated the rapid and strong antiviral activities of AsIFN-γ against RGNNV in black seabream.
Keywords
ISGF3NLSGATA3NF-κBRdRpISGIFN-gammaJAK-STATISREIRFhour post infectionOCT1calcium/calmodulin kinase IITCID50Janus kinase-signal transducers and activators of transcriptionRGNNVGATA-binding protein 3IFN-stimulated gene factor 3PBSCaMKIIqRT-PCRhpiAP-1ORFPI3-K50% tissue culture infective doseISGsRNA-dependent RNA polymeraseinterferonIFNbase pairBlack seabreamnuclear localization signalinterferon regulatory factorinterferon-stimulated response elementnuclear factor kBopen reading framephosphate buffer solutionUTR یا untranslated regions untranslated regionquantitative real time polymerase chain reactionactivator protein 1mitogen activated protein kinasePoly I:Cpolyinosinic-polycytidylic acidInterferon-stimulated genesMAP kinaseGas
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Authors
Yangxi Xiang, Wei Liu, Peng Jia, Yunlong Li, Yilin Jin, Limin Chen, Jing Zhang, Kuntong Jia, Meisheng Yi,