Characterization and Transcript Expression Analyses of Atlantic Cod Viperin
| Autor: | Matthew L. Rise, Khalil Eslamloo, Sabrina M. Inkpen, Mani Larijani, Atefeh Ghorbani, Xi Xue |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Fish Proteins
lcsh:Immunologic diseases. Allergy 0301 basic medicine DNA Complementary Transcription Genetic Immunology dsRNA rsad2 03 medical and health sciences 0302 clinical medicine Complementary DNA Gene expression teleost ISGs Animals Immunology and Allergy Gadus Gene Original Research biology Gene Expression Profiling Macrophages Exons biology.organism_classification Protein kinase R Cell biology qPCR RNA silencing Poly I-C 030104 developmental biology Gadus morhua inhibition of antiviral responses Viperin RNA Interferons lcsh:RC581-607 Atlantic cod Signal Transduction 030215 immunology |
| Zdroj: | Frontiers in Immunology, Vol 10 (2019) Frontiers in Immunology |
| ISSN: | 1664-3224 |
| Popis: | Viperin is a key antiviral effector in immune responses of vertebrates including the Atlantic cod (Gadus morhua). Using cloning, sequencing and gene expression analyses, we characterized the Atlantic cod viperin at the nucleotide and hypothetical amino acid levels, and its regulating factors were investigated. Atlantic cod viperin cDNA is 1,342 bp long, and its predicted protein contains 347 amino acids. Using in silico analyses, we showed that Atlantic cod viperin is composed of 5 exons, as in other vertebrate orthologs. In addition, the radical SAM domain and C-terminal sequences of the predicted Viperin protein are highly conserved among various species. As expected, Atlantic cod Viperin was most closely related to other teleost orthologs. Using computational modeling, we show that the Atlantic cod Viperin forms similar overall protein architecture compared to mammalian Viperins. qPCR revealed that viperin is a weakly expressed transcript during embryonic development of Atlantic cod. In adults, the highest constitutive expression of viperin transcript was found in blood compared with 18 other tissues. Using isolated macrophages and synthetic dsRNA (pIC) stimulation, we tested various immune inhibitors to determine the possible regulating pathways of Atlantic cod viperin. Atlantic cod viperin showed a comparable pIC induction to other well-known antiviral genes (e.g., interferon gamma and interferon-stimulated gene 15-1) in response to various immune inhibitors. The pIC induction of Atlantic cod viperin was significantly inhibited with 2-Aminopurine, Chloroquine, SB202190, and Ruxolitinib. Therefore, endosomal-TLR-mediated pIC recognition and signal transducers (i.e., PKR and p38 MAPK) downstream of the TLR-dependent pathway may activate the gene expression response of Atlantic cod viperin. Also, these results suggest that antiviral responses of Atlantic cod viperin may be transcriptionally regulated through the interferon-activated pathway. |
| Databáze: | OpenAIRE |
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