Department of Entomology, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran.
Australian Infectious Disease Research Centre, School of Biological Sciences, The University of Queensland, Brisbane, QLD, Australia.
Insect Biochem Mol Biol. 2019 Sep;112:103202. doi: 10.1016/j.ibmb.2019.103202. Epub 2019 Aug 15.
The microRNA (miRNA) pathway is an epigenetic mechanism that plays important roles in various biological processes including host-virus interactions by regulating gene expression of the host and/or the virus. Previously, we showed that the cellular microRNAome in Spodoptera frugiperda (Sf9) cells is modulated following Autographa californica multiple nucleopolyhedrovirus (AcMNPV) infection suggesting that miRNAs may contribute in the cellular antiviral immunity. Here, we investigated the role of core components of the miRNA pathway in Sf9-AcMNPV interaction. Gene expression analyses showed that the expression levels of Dicer-1 (Dcr1), Argonaute-1 (Ago1) and Exportin-5 (Exp5) increased following AcMNPV infection particularly at 16 h post infection (hpi). Ran expression levels, however, decreased in response to virus infection. The expression levels of cellular miRNAs, miR-184 and let-7, also diminished at the post infection times further confirming differential expression of the cellular miRNAs following AcMNPV infection. To determine the role of the miRNA pathway in the interaction, we silenced key genes in the pathway using specific dsRNAs. RNAi of Dcr1, Ago1 and Ran enhanced viral DNA replication and reduced the abundance of miR-184 and let-7 underscoring the importance of the miRNA pathway in antiviral immunity in Sf9 cells. Suppression of the miRNA pathway in mock and infected cells had no effect on Ran expression levels suggesting miRNA-independent downregulation of this gene after virus infection. In conclusion, our results suggest the antiviral role of the miRNA pathway in Sf9 cells against AcMNPV. To modulate this immune response, AcMNPV represses host miRNAs likely through downregulation of Ran to enhance its replication in the host cells.
微 RNA(miRNA)途径是一种表观遗传机制,通过调节宿主和/或病毒的基因表达,在包括宿主-病毒相互作用在内的各种生物学过程中发挥重要作用。之前,我们表明,在感染 Autographa californica 多角体病毒(AcMNPV)后,Spodoptera frugiperda( Sf9)细胞中的细胞 microRNAome 会发生变化,这表明 miRNA 可能有助于细胞抗病毒免疫。在这里,我们研究了 miRNA 途径的核心成分在 Sf9-AcMNPV 相互作用中的作用。基因表达分析表明,Dicer-1(Dcr1)、Argonaute-1(Ago1)和 Exportin-5(Exp5)的表达水平在 AcMNPV 感染后增加,特别是在感染后 16 小时(hpi)。然而,Ran 的表达水平因病毒感染而下降。细胞 miRNA,miR-184 和 let-7 的表达水平也在感染后时间进一步减少,进一步证实了 AcMNPV 感染后细胞 miRNA 的差异表达。为了确定 miRNA 途径在相互作用中的作用,我们使用特异性 dsRNA 沉默了途径中的关键基因。Dcr1、Ago1 和 Ran 的 RNAi 增强了病毒 DNA 复制,并减少了 miR-184 和 let-7 的丰度,这突出了 miRNA 途径在 Sf9 细胞抗病毒免疫中的重要性。在模拟和感染细胞中抑制 miRNA 途径对 Ran 表达水平没有影响,这表明在病毒感染后,该基因的下调是 miRNA 非依赖性的。总之,我们的研究结果表明,miRNA 途径在 Sf9 细胞中对 AcMNPV 具有抗病毒作用。为了调节这种免疫反应,AcMNPV 通过下调 Ran 来抑制宿主 miRNA,从而增强其在宿主细胞中的复制。