Molecular cloning, functional characterization and antiviral activity of porcine DDX3X
Molecular cloning, functional characterization and antiviral activity of porcine DDX3X
复制标题
猪DDX3X的分子克隆、功能表征和抗病毒活性
DOI：
10.1016/j.bbrc.2013.12.098
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发表时间：
2014-01-24
影响因子：
3.1
通讯作者：
Fang, Liurong
中科院分区：
文献类型：
--
作者：
Chen, Quangang;Liu, Qin;Fang, Liurong
人DDX 3X是一种新发现的DEAD盒RNA解旋酶。除了真核基因表达调控的参与，人DDX 3X最近已被证明是先天免疫信号传导途径中的关键分子，并有助于I型干扰素（IFN）诱导。本研究通过RT-PCR从PK-15细胞中克隆了猪DDX 3X基因，并对其调控IFN-β的功能进行了研究。推测的猪DDX 3X ORF编码662个氨基酸，具有几个保守的基序。序列比对结果表明，猪DDX 3X与马（96.7%）、小鼠（97.6%）、牛（98.5%）、狗（98.6%）和人（98.9%）的DDX 3X在氨基酸水平上具有较高的同源性。猪DDX 3X的异位表达显著激活IFN-β表达，而猪DDX 3X的敲低抑制dsRNA或仙台病毒（SeV）诱导的IFN-β。此外，猪DDX 3X与IPS-1、TBK 1和IKK β共定位，并增强这些分子诱导的IFN-β启动子激活。我们还研究了猪DDX 3X在猪繁殖与呼吸综合征病毒（PRRSV）感染过程中的作用，发现DDX 3X的过表达显著抑制了PRRSV的复制，表明DDX 3X是一种潜在的抗病毒剂。(C)2013 Elsevier Inc. All rights reserved.
Human DDX3X is a newly discovered DEAD-box RNA helicase. In addition to involvement of eukaryotic gene expression regulation, human DDX3X has recently been demonstrated to be a critical molecule in innate immune signaling pathways and to contribute to type I interferon (IFN) induction. In the present study, porcine DDX3X was cloned by RT-PCR from PK-15 cells and its function in regulating IFN-beta was characterized. The putative porcine DDX3X ORF encodes 662 amino acids possessing several conserved motifs. Sequence alignments indicated that porcine DDX3X has high identity at the amino acid level to those of horse (96.7%), mouse (97.6%), cattle (98.5%), dog (98.6%) and human (98.9%). Ectopic expression of porcine DDX3X significantly activated IFN-beta expression, whereas knockdown of porcine DDX3X inhibited dsRNA- or Sendai virus (SeV)-induced IFN-beta. Furthermore, porcine DDX3X co-localized with IPS-1, TBK1 and IKK epsilon, and enhanced IFN-beta promoter activation induced by these molecules. We also investigated the role of porcine DDX3X during porcine reproductive and respiratory syndrome virus (PRRSV) infection and found that overexpression of DDX3X significantly inhibited PRRSV replication, indicating that DDX3X is a potential antiviral agent. (C) 2013 Elsevier Inc. All rights reserved.