NF-κB Signaling Differentially Regulates Influenza Virus RNA Synthesis
NF-κB Signaling Differentially Regulates Influenza Virus RNA Synthesis
复制标题
DOI：
10.1128/jvi.00909-08
复制
发表时间：
2008-10-01
影响因子：
5.4
通讯作者：
Liang, Yuying
中科院分区：
文献类型：
--
作者：
Kumar, Naveen;Xin, Zhong-tao;Liang, Yuying
NF-kB信号通路先前已被证明是有效的甲型流感病毒复制所需的，尽管分子机制还不清楚。在这项研究中，我们通过使用两种已知的NF-κ B抑制剂和多种流感病毒特异性检测，确定了受NF-κ B B信号传导影响的流感病毒生命周期的特定步骤。时程实验的结果表明，NF-κ B抑制剂Bay 11 -7082和吡咯烷二硫代氨基甲酸铵抑制病毒感染的早期posttry步骤，但它们似乎并不影响病毒核糖核蛋白复合物的核质运输。相反，我们发现，流感病毒基因组RNA（vRNA）的水平，但不是相应的cRNA或mRNA，特异性地减少病毒感染细胞中的抑制剂，表明NF-κ B信号密切参与vRNA的合成。此外，我们在报道基因分析中发现NF-κ B抑制剂特异性地减少了流感病毒RNA从cRNA启动子的转录，但不减少从vRNA启动子的转录，这一结果与从病毒感染细胞获得的数据一致。p65 NF-κ B分子的过表达不仅可以消除抑制作用，而且可以激活流感病毒RNA从cRNA启动子的转录。最后，使用p65特异性小干扰RNA，我们已经表明，p65敲低降低流感病毒复制和vRNA合成的水平。综上所述，我们首次提供证据表明NF-κ B B宿主信号通路可以差异调节流感病毒RNA合成，这也可能为理解其他RNA病毒对RNA合成的宿主调节提供一些新的视角。
The NF-kB signaling pathway has previously been shown to be required for efficient influenza A virus replication, although the molecular mechanism is not well understood. In this study, we identified a specific step of the influenza virus life cycle that is influenced by NF-kappa B signaling by using two known NF-kappa B inhibitors and a variety of influenza virus-specific assays. The results of time course experiments suggest that the NF-kappa B inhibitors Bay11-7082 and ammonium pyrrolidinedithiocarbamate inhibited an early postentry step of viral infection, but they did not appear to affect the nucleocytoplasmic trafficking of the viral ribonucleoprotein complex. Instead, we found that the levels of influenza virus genomic RNA (vRNA), but not the corresponding cRNA or mRNA, were specifically reduced by the inhibitors in virus-infected cells, indicating that NF-kappa B signaling is intimately involved in the vRNA synthesis. Furthermore, we showed that the NF-kappa B inhibitors specifically diminished influenza virus RNA transcription from the cRNA promoter but not from the vRNA promoter in a reporter assay, a result which is consistent with data obtained from virus-infected cells. The overexpression of the p65 NF-kappa B molecule could not only eliminate the inhibition but also activate influenza virus RNA transcription from the cRNA promoter. Finally, using p65-specific small interfering RNA, we have shown that p65 knockdown reduced the levels of influenza virus replication and vRNA synthesis. In summary, we have provided evidence showing, for the first time, that the NF-kappa B host signaling pathway can differentially regulate influenza virus RNA synthesis, which may also offer some new perspectives into understanding the host regulation of RNA synthesis by other RNA viruses.