Bi Ke-Ran, Han Kai-Kai, Liu Qing-Tao, Zhao Dong-Min, Huang Xin-Mei, Liu Yu-Zhuo, Yang Jing, Li Yin
Key Laboratory of Veterinary Biological Engineering and Technology, National Center for Engineering Research of Veterinary Bio-products, Institute of Veterinary Medicine, Ministry of Agriculture, Jiangsu Academy of Agricultural Sciences, Nanjing, China; Jiangsu Key Laboratory for Marine Biotechnology, Co-Innovation Center of Jiangsu Marine Bio-industry Technology, Huaihai Institute of Technology, Lian Yungang, China.
Key Laboratory of Veterinary Biological Engineering and Technology, National Center for Engineering Research of Veterinary Bio-products, Institute of Veterinary Medicine, Ministry of Agriculture, Jiangsu Academy of Agricultural Sciences, Nanjing, China.
Gene. 2017 Sep 20;629:43-51. doi: 10.1016/j.gene.2017.07.067. Epub 2017 Jul 25.
2'-5'-Oligoadenylate synthetase-like protein (OASL) is an interferon-inducible antiviral protein that exerts antiviral effects through the RNase L- or retinoic acid-inducible gene I (RIG-I)-dependent signalling pathway. In this study, we identified and cloned the OASL gene (named duOASL) from healthy adult Cherry Valley ducks. Full-length duOASL cDNA (1630bp) encoded a 504-amino acid polypeptide containing three conserved domains, namely, nucleotidyltransferase domain, 2'-5'-oligoadenylate synthetase domain, and two ubiquitin-like repeats. DuOASL mRNA expression was quantified by performing quantitative reverse transcription-PCR (qRT-PCR). Results of qRT-PCR showed that duOASL was broadly expressed in all examined tissues, with the highest mRNA expression in the large intestine. Antiviral activity of duOASL was measured by determining its effect on Duck Tembusu virus (DTMUV) replication in vitro. We found that duOASL overexpression slightly inhibited DTMUV replication, whereas duOASL knockdown by using a specific small interfering RNA increased DTMUV replication in DF-1 cells. Thus, we successfully cloned and characterized the antiviral protein duOASL from Cherry Valley ducks and found that it exerted antiviral effects against DTMUV. These results provide a solid foundation for performing further studies to determine the mechanism underlying the antiviral effect of duOASL at the cellular level.
2'-5'-寡腺苷酸合成酶样蛋白（OASL）是一种干扰素诱导的抗病毒蛋白，它通过核糖核酸酶L或视黄酸诱导基因I（RIG-I）依赖性信号通路发挥抗病毒作用。在本研究中，我们从健康成年樱桃谷鸭中鉴定并克隆了OASL基因（命名为duOASL）。全长duOASL cDNA（1630bp）编码一个504个氨基酸的多肽，包含三个保守结构域，即核苷酸转移酶结构域、2'-5'-寡腺苷酸合成酶结构域和两个泛素样重复序列。通过定量逆转录PCR（qRT-PCR）对duOASL mRNA表达进行定量。qRT-PCR结果显示，duOASL在所有检测组织中广泛表达，并在大肠中具有最高的mRNA表达水平。通过测定duOASL对鸭坦布苏病毒（DTMUV）体外复制的影响来检测其抗病毒活性。我们发现，duOASL过表达略微抑制DTMUV复制，而使用特异性小干扰RNA敲低duOASL则增加了DF-1细胞中DTMUV的复制。因此，我们成功地从樱桃谷鸭中克隆并鉴定了抗病毒蛋白duOASL，并发现它对DTMUV具有抗病毒作用。这些结果为进一步研究duOASL在细胞水平上抗病毒作用的机制奠定了坚实的基础。
