Host factor Rab11a is critical for efficient assembly of influenza A virus genomic segments.
Host factor Rab11a is critical for efficient assembly of influenza A virus genomic segments.
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DOI：
10.1371/journal.ppat.1009517
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发表时间：
2021-05
期刊：
影响因子：
6.7
通讯作者：
Manicassamy B
中科院分区：
文献类型：
--
作者：
Han J;Ganti K;Sali VK;Twigg C;Zhang Y;Manivasagam S;Liang CY;Vogel OA;Huang I;Emmanuel SN;Plung J;Radoshevich L;Perez JT;Lowen AC;Manicassamy B
甲型流感病毒选择性地将8个不同的病毒核糖核蛋白复合体(VRNP)包装到每个病毒粒子中；然而，宿主因素在基因组组装中的作用尚不完全清楚。为了评价细胞因子在基因组组装中的意义，我们构建了一个在NP基因中带有四半胱氨酸标签的报告病毒(NP-TC病毒)，并在荧光显微镜下评估了vRNP随细胞成分的定位动态。在早期，vRNP复合体优先输出到MTOC；随后，vRNP结合在细胞因子Rab11a阳性的小泡上，形成不同的vRNP束，在微管网络上运输到质膜。然而，在Rab11a缺失的细胞中，vRNP束在细胞质中较小，不同vRNP片段之间的共定位较少。此外，Rab11a缺乏增加了非感染性颗粒的产生，具有更高的RNA拷贝数与PFU比率，表明特定基因组组装存在缺陷。这些结果表明，Rab11a+囊泡是vRNP复合体聚集的枢纽，并通过vRNP：vRNP相互作用实现特定的基因组组装，揭示了Rab11a作为甲型流感病毒基因组组装的关键宿主因子的重要性。甲型流感病毒(IAV)基因组由8个不同的RNA片段组成。目前尚不清楚这8个单独的RNA片段是如何组装在一起形成具有感染性的病毒颗粒的。我们的研究表明，Rab11a+囊泡是聚集和组装8个单独的病毒RNA片段的平台，这些片段是形成感染性病毒颗粒所需的。然而，在缺乏Rab11a的细胞中，病毒RNA片段无法聚集在一起，导致缺陷病毒颗粒的产生增加，这可能是由于病毒RNA片段的错误组装。因此，我们的研究揭示了Rab11a在流感病毒基因组组装和传染性病毒颗粒生产中的重要作用。
It is well documented that influenza A viruses selectively package 8 distinct viral ribonucleoprotein complexes (vRNPs) into each virion; however, the role of host factors in genome assembly is not completely understood. To evaluate the significance of cellular factors in genome assembly, we generated a reporter virus carrying a tetracysteine tag in the NP gene (NP-Tc virus) and assessed the dynamics of vRNP localization with cellular components by fluorescence microscopy. At early time points, vRNP complexes were preferentially exported to the MTOC; subsequently, vRNPs associated on vesicles positive for cellular factor Rab11a and formed distinct vRNP bundles that trafficked to the plasma membrane on microtubule networks. In Rab11a deficient cells, however, vRNP bundles were smaller in the cytoplasm with less co-localization between different vRNP segments. Furthermore, Rab11a deficiency increased the production of non-infectious particles with higher RNA copy number to PFU ratios, indicative of defects in specific genome assembly. These results indicate that Rab11a+ vesicles serve as hubs for the congregation of vRNP complexes and enable specific genome assembly through vRNP:vRNP interactions, revealing the importance of Rab11a as a critical host factor for influenza A virus genome assembly. The influenza A virus (IAV) genome is composed of 8 distinct RNA segments. It has remained unclear how these 8 individual RNA segments are assembled together to form infectious virus particles. Our study shows that Rab11a+ vesicles serve as platforms for the congregation and assembly of 8 individual viral RNA segments needed to form infectious virus particles. However, in cells lacking Rab11a, viral RNA segments fail to congregate together, resulting in increased production of defective virus particles, likely due to misassembling of viral RNA segments. Thus, our study reveals the important role for Rab11a in influenza virus genome assembly and production of infectious virus particles.
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作者：
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影响因子：
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作者：
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作者：
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作者：
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DOI：
10.1038/nrmicro3367
发表时间：
2015-01
期刊：
Nature reviews. Microbiology
影响因子：
--
作者：
Eisfeld AJ;Neumann G;Kawaoka Y
通讯作者：
Kawaoka Y