Department of Ocean Sciences, Memorial University of Newfoundland, St. John's, NL, Canada.
Division of Biomedical Sciences, Faculty of Medicine, Memorial University of Newfoundland, St. John's, NL, Canada.
Front Immunol. 2019 Mar 6;10:311. doi: 10.3389/fimmu.2019.00311. eCollection 2019.
Viperin is a key antiviral effector in immune responses of vertebrates including the Atlantic cod (). Using cloning, sequencing and gene expression analyses, we characterized the Atlantic cod at the nucleotide and hypothetical amino acid levels, and its regulating factors were investigated. Atlantic cod cDNA is 1,342 bp long, and its predicted protein contains 347 amino acids. Using analyses, we showed that Atlantic cod 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 is a weakly expressed transcript during embryonic development of Atlantic cod. In adults, the highest constitutive expression of 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 . Atlantic cod showed a comparable pIC induction to other well-known antiviral genes (e.g., and ) in response to various immune inhibitors. The pIC induction of Atlantic cod 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 . Also, these results suggest that antiviral responses of Atlantic cod may be transcriptionally regulated through the interferon-activated pathway.
Viperin 是脊椎动物（包括大西洋鳕鱼）免疫反应中的一种关键抗病毒效应因子。通过克隆、测序和基因表达分析，我们在核苷酸和假设的氨基酸水平上对大西洋鳕鱼 进行了特征描述，并研究了其调节因子。大西洋鳕鱼 cDNA 长 1342bp，其预测蛋白包含 347 个氨基酸。通过 分析，我们表明大西洋鳕鱼 由 5 个外显子组成，与其他脊椎动物同源物相同。此外，预测的 Viperin 蛋白的激进 SAM 结构域和 C 末端序列在各种物种中高度保守。不出所料，大西洋鳕鱼 Viperin 与其他硬骨鱼的同源物最为密切相关。通过计算建模，我们表明大西洋鳕鱼 Viperin 与哺乳动物 Viperins 形成相似的整体蛋白质结构。qPCR 显示，在大西洋鳕鱼胚胎发育过程中， 是一种低表达的转录本。在成鱼中，与 18 种其他组织相比，血液中 的转录本表达量最高。使用分离的巨噬细胞和合成 dsRNA（pIC）刺激，我们测试了各种免疫抑制剂，以确定大西洋鳕鱼 的可能调节途径。与其他著名的抗病毒基因（如 和 ）一样，大西洋鳕鱼 在对各种免疫抑制剂的反应中表现出相当的 pIC 诱导。用 2-氨基嘌呤、氯喹、SB202190 和鲁索替尼处理后，大西洋鳕鱼 的 pIC 诱导显著受到抑制。因此，内体 TLR 介导的 pIC 识别和 TLR 依赖性途径下游的信号转导物（即 PKR 和 p38 MAPK）可能激活大西洋鳕鱼 的基因表达反应。此外，这些结果表明，大西洋鳕鱼 的抗病毒反应可能通过干扰素激活途径进行转录调控。
