A Rab11-and Microtubule-Dependent Mechanism for Cytoplasmic Transport of Influenza A Virus Viral RNA
A Rab11-and Microtubule-Dependent Mechanism for Cytoplasmic Transport of Influenza A Virus Viral RNA
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DOI：
10.1128/jvi.02606-10
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发表时间：
2011-05-01
影响因子：
5.4
通讯作者：
Digard, Paul
中科院分区：
文献类型：
--
作者：
Amorim, Maria Joao;Bruce, Emily A.;Digard, Paul
甲型流感病毒的病毒RNA（vRNA）基因组在细胞核中复制，作为核糖核蛋白（RNP）输出到细胞质，并通过不确定的方式运输到质膜。使用荧光原位杂交检测vRNA以及荧光标记的RNP的活细胞成像，我们表明，vRNA细胞质运输的早期事件涉及在多种细胞类型和病毒株的微管组织中心附近的积累。在这里，RNP与Rab11共定位，Rab11是一种中心粒周围再循环内体标记物。胞质RNP本地化受到干扰抑制剂的囊泡贩运，微管，或短干扰RNA介导的Rab11的耗尽。活细胞中的绿色荧光蛋白（GFP）标记的RNP表现出快速、双向和跳跃式运动，这是基于微管的运输的特征，并且还与荧光Rab11共同运输。共沉淀实验表明RNP和GTP结合形式的Rab11之间的相互作用，可能通过PB2亚基的聚合酶介导。我们建议，流感病毒RNP路由从细胞核到pericentriolar再循环内体（RE），在那里他们访问Rab11依赖的囊泡运输途径的细胞周边。
The viral RNA (vRNA) genome of influenza A virus is replicated in the nucleus, exported to the cytoplasm as ribonucleoproteins (RNPs), and trafficked to the plasma membrane through uncertain means. Using fluorescent in situ hybridization to detect vRNA as well as the live cell imaging of fluorescently labeled RNPs, we show that an early event in vRNA cytoplasmic trafficking involves accumulation near the microtubule organizing center in multiple cell types and viral strains. Here, RNPs colocalized with Rab11, a pericentriolar recycling endosome marker. Cytoplasmic RNP localization was perturbed by inhibitors of vesicular trafficking, microtubules, or the short interfering RNA-mediated depletion of Rab11. Green fluorescent protein (GFP)-tagged RNPs in living cells demonstrated rapid, bidirectional, and saltatory movement, which is characteristic of microtubule-based transport, and also cotrafficked with fluorescent Rab11. Coprecipitation experiments showed an interaction between RNPs and the GTP-bound form of Rab11, potentially mediated via the PB2 subunit of the polymerase. We propose that influenza virus RNPs are routed from the nucleus to the pericentriolar recycling endosome (RE), where they access a Rab11-dependent vesicular transport pathway to the cell periphery.