对甲型流感病毒mRNA在活细胞中的转运进行成像与表征。
Imaging and characterizing influenza A virus mRNA transport in living cells.
作者信息
Wang Wei, Cui Zong-Qiang, Han Han, Zhang Zhi-Ping, Wei Hong-Ping, Zhou Ya-Feng, Chen Ze, Zhang Xian-En
机构信息
State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, China.
出版信息
Nucleic Acids Res. 2008 Sep;36(15):4913-28. doi: 10.1093/nar/gkn475. Epub 2008 Jul 24.
The mechanisms of influenza A virus mRNA intracellular transport are still not clearly understood. Here, we visualized the distribution and transport of influenza A virus mRNA in living cells using molecular beacon (MB) technology. Confocal-FRAP measurements determined that the transport of influenza A virus intronless mRNA, in both nucleus and cytoplasm, was energy dependent, being similar to that of Poly(A)(+) RNA. Drug inhibition studies in living cells revealed that the export of influenza A virus mRNA is independent of the CRM1 pathway, while the function of RNA polymerase II (RNAP-II) may be needed. In addition, viral NS1 protein and cellular TAP protein were found associated with influenza A virus mRNA in the cell nucleus. These findings characterize influenza A virus mRNA transport in living cells and suggest that influenza A virus mRNA may be exported from the nucleus by the cellular TAP/p15 pathway with NS1 protein and RNAP-II participation.
甲型流感病毒mRNA的细胞内运输机制仍未完全清楚。在此，我们使用分子信标（MB）技术观察了甲型流感病毒mRNA在活细胞中的分布和运输情况。共聚焦荧光恢复后光漂白（Confocal-FRAP）测量结果表明，甲型流感病毒无内含子mRNA在细胞核和细胞质中的运输均依赖能量，这与聚腺苷酸（Poly(A)(+)）RNA的运输情况相似。对活细胞进行的药物抑制研究显示，甲型流感病毒mRNA的输出不依赖CRM1途径，但可能需要RNA聚合酶II（RNAP-II）发挥作用。此外，在细胞核中发现病毒NS1蛋白和细胞TAP蛋白与甲型流感病毒mRNA相关联。这些发现明确了甲型流感病毒mRNA在活细胞中的运输特征，并表明甲型流感病毒mRNA可能在NS1蛋白和RNAP-II的参与下，通过细胞TAP/p15途径从细胞核输出。
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