West nile virus-induced interferon production is mediated by the double-stranded RNA-Dependent protein kinase PKR
West nile virus-induced interferon production is mediated by the double-stranded RNA-Dependent protein kinase PKR
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DOI：
10.1128/jvi.00446-07
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发表时间：
2007-10-01
影响因子：
5.4
通讯作者：
Mason, Peter W.
中科院分区：
文献类型：
--
作者：
Gilfoy, Felicia D.;Mason, Peter W.
细胞携带各种分子，称为病原体识别受体(PRRs)，能够感知入侵的病原体。PRRs与病毒化合物的相互作用激发了一条信号通路(S)，导致包括I型干扰素(干扰素)在内的基因激活，这些基因对有效的抗病毒反应至关重要。在这里，我们证明了双链RNA(DsRNA)依赖的蛋白激酶PKR在经dsRNA模拟物Poly(I：C)处理的细胞中作为PRR发挥功能，在西尼罗河病毒(WNV)感染的细胞中作为PRR发挥作用。从人和小鼠细胞中都获得了PKR作为PRR的作用的证据。在小鼠胚胎成纤维细胞(MEF)中，我们证明了PKR基因敲除、使用小干扰RNA(SiRNA)抑制PKR基因转录后的基因沉默以及化学抑制PKR功能都干扰了西尼罗河病毒感染后干扰素的合成。在三种不同的人类细胞系中，siRNA敲除和化学抑制PKR阻断了西尼罗河病毒诱导的干扰素的合成。使用相同的方法，我们证明了PKR不是仙台病毒诱导的干扰素合成所必需的，这表明PKR对于识别西尼罗河病毒感染特别重要。综上所述，我们的数据表明，PKR可以作为识别西尼罗河病毒感染的PRR。
Cells carry a variety of molecules, referred to as pathogen recognition receptors (PRRs), which are able to sense invading pathogens. Interaction of PRRs with viral compounds instigates a signaling pathway(s), resulting in the activation of genes, including those for type I interferon (IFN), which are critical for an effective antiviral response. Here we demonstrate that the double-stranded RNA (dsRNA)-dependent protein kinase PKR, which has been shown to function as a PRR in cells treated with the dsRNA mimetic poly(I:C), serves as a PRR in West Nile virus (WNV)-infected cells. Evidence for PKR's role as a PRR was obtained from both human and murine cells. Using mouse embryonic fibroblasts (MEFs), we demonstrated that PKR gene knockout, posttranscriptional gene silencing of PKR mRNA using small interfering RNA (siRNA), and chemical inhibition of PKR function all interfered with IFN synthesis following WNV infection. In three different human cell lines, siRNA knockdown and chemical inhibition of PKR blocked WNV-induced IFN synthesis. Using the same approaches, we demonstrated that PKR was not necessary for Sendai virus-induced IFN synthesis, suggesting that PKR is particularly important for recognition of WNV infection. Taken together, our data suggest that PKR could serve as a PRR for recognition of WNV infection.