RAB11A Is Essential for Transport of the Influenza Virus Genome to the Plasma Membrane
RAB11A Is Essential for Transport of the Influenza Virus Genome to the Plasma Membrane
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DOI：
10.1128/jvi.00378-11
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发表时间：
2011-07-01
影响因子：
5.4
通讯作者：
Kawaoka, Yoshihiro
中科院分区：
文献类型：
--
作者：
Eisfeld, Amie J.;Kawakami, Eiryo;Kawaoka, Yoshihiro
甲型流感病毒组装是一个复杂的过程，需要参与将病毒糖蛋白、基质蛋白和病毒基因组（并入病毒核糖核蛋白（vRNP）复合物）转运至病毒体形成的质膜位点的途径交叉。在这些病毒体组分中，vRNP递送的机制是最不完全了解的。在这里，我们揭示了细胞Rab11 GTP酶亚型，RAB11A，和vRNP之间的功能关系，并表明RAB11A是必不可少的适当vRNP运输到质膜。使用单克隆抗体，识别核蛋白的形式vRNP的免疫荧光为基础的测定，我们证明之间的关联RAB11 A和vRNP在vRNP胞质转运的所有阶段。通过小干扰RNA（siRNA）处理或通过显性负性或组成型活性蛋白的过表达破坏RAB11A功能，导致异常vRNP细胞内蓄积，保留在核周区域，并缺乏质膜蓄积。RAB11A和vRNP之间的复合物形成进一步建立生化。我们的研究结果揭示了一个关键的宿主因子，对流感病毒基因组的传递有重要贡献，并揭示了RAB11A在核糖核蛋白运输中的潜在作用。
Influenza A virus assembly is a complex process that requires the intersection of pathways involved in transporting viral glycoproteins, the matrix protein, and viral genomes, incorporated in the viral ribonucleoprotein (vRNP) complex, to plasma membrane sites of virion formation. Among these virion components, the mechanism of vRNP delivery is the most incompletely understood. Here, we reveal a functional relationship between the cellular Rab11 GTPase isoform, RAB11A, and vRNPs and show that RAB11A is indispensable for proper vRNP transport to the plasma membrane. Using an immunofluorescence-based assay with a monoclonal antibody that recognizes nucleoprotein in the form of vRNP, we demonstrate association between RAB11A and vRNPs at all stages of vRNP cytoplasmic transport. Abrogation of RAB11A expression through small interfering RNA (siRNA) treatment or disruption of RAB11A function by overexpression of dominant negative or constitutively active proteins caused aberrant vRNP intracellular accumulation, retention in the perinuclear region, and lack of accumulation at the plasma membrane. Complex formation between RAB11A and vRNPs was further established biochemically. Our results uncover a critical host factor with an essential contribution to influenza virus genome delivery and reveal a potential role for RAB11A in the transport of ribonucleoprotein cargo.