Characterization and Transcript Expression Analyses of Atlantic Cod Viperin
Characterization and Transcript Expression Analyses of Atlantic Cod Viperin
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DOI：
10.3389/fimmu.2019.00311
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发表时间：
2019-03-06
影响因子：
7.3
通讯作者：
Rise, Matthew L.
中科院分区：
文献类型：
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作者：
Eslamloo, Khalil;Ghorbani, Atefeh;Rise, Matthew L.
蝰蛇蛋白是脊椎动物包括大西洋鳕鱼（Gadus morhua）的免疫应答中的关键抗病毒效应物。通过克隆、测序和基因表达分析，我们在核苷酸和假定氨基酸水平上对大西洋鳕鱼蝰蛇蛋白进行了表征，并对其调控因子进行了研究。大西洋鳕鱼蝰蛇蛋白cDNA全长1，342 bp，预测的蛋白质由347个氨基酸组成。使用计算机分析，我们表明，大西洋鳕鱼蝰蛇蛋白是由5个外显子，在其他脊椎动物的直系同源。此外，预测的Viperin蛋白的自由基SAM结构域和C-末端序列在各种物种中高度保守。正如预期的那样，大西洋鳕鱼蝰蛇蛋白与其他硬骨鱼类直系同源物关系最密切。使用计算模型，我们表明，大西洋鳕鱼蝰蛇蛋白形成类似的整体蛋白质结构相比，哺乳动物蝰蛇。qPCR结果表明，viperin在大西洋鳕鱼胚胎发育过程中是一个弱表达的转录本。在成年人中，与其他18种组织相比，在血液中发现了最高的蝰蛇蛋白转录本组成型表达。使用分离的巨噬细胞和合成的dsRNA（pIC）刺激，我们测试了各种免疫抑制剂，以确定可能的调节途径大西洋鳕鱼蝰蛇蛋白。大西洋鳕鱼蝰蛇蛋白显示出与其他公知的抗病毒基因相当的pIC诱导（e.例如，在一个实施例中，干扰素γ和干扰素刺激基因15-1）对各种免疫抑制剂的应答。2-氨基嘌呤、氯喹、SB 202190和Ruxolitinib显著抑制大西洋鳕鱼蝰蛇蛋白的pIC诱导。因此，内体TLR介导的pIC识别和信号转导（即，例如，PKR和p38 MAPK）的TLR依赖性途径下游可能激活大西洋鳕鱼蝰蛇蛋白的基因表达反应。此外，这些结果表明，大西洋鳕鱼蝰蛇蛋白的抗病毒反应可能是通过干扰素激活途径转录调节。
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.