Host Retromer Protein Sorting Nexin 2 Interacts with Human Respiratory Syncytial Virus Structural Proteins and is Required for Efficient Viral Production.
Host Retromer Protein Sorting Nexin 2 Interacts with Human Respiratory Syncytial Virus Structural Proteins and is Required for Efficient Viral Production.
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DOI：
10.1128/mbio.01869-20
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发表时间：
2020-09-29
期刊：
影响因子：
6.4
通讯作者：
Arruda E
中科院分区：
文献类型：
--
作者：
Cardoso RS;Tavares LA;Jesus BLS;Criado MF;de Carvalho AN;Souza JP;Bedi S;de Souza MM;Silva ML;Lanfredi GP;Vitti BC;Scudero OB;Faça VM;Ono A;Ventura AM;daSilva LLP;Arruda E
本研究提供了新的知识，以了解HRSV组装提供的证据表明，非糖基化的结构蛋白M和N与分泌途径的元素相互作用，揭示其细胞内交通。据我们所知，目前的贡献是重要的，因为缺乏有关HRSV非糖基化蛋白质的交通研究，特别是通过指出SNX2，一个retromer组件，在HRSV组装过程中的参与。人呼吸道合胞病毒（HRSV）包膜糖蛋白通过分泌途径运输到组装位点，而非糖基化蛋白M和N存在于HRSV包涵体中，但必须到达质膜，在那里HRSV组装发生。关于非糖基化的HRSV蛋白如何到达组装位点知之甚少。在这里，我们表明，HRSV M和N蛋白部分共定位与高尔基体标记giantin，和糖基化的F和nonglycosylated N蛋白紧密位于反式高尔基体，这表明他们的相互作用，在该车厢。Brefeldin A损害HRSV F和N蛋白的运输和包涵体大小，表明高尔基体对糖基化和非糖基化HRSV蛋白运输都很重要。HRSV N和M蛋白与分选连接蛋白2（SNX2）共定位并相互作用，SNX2是一种在管状结构中形成内体的逆转录组分。糖基化F和非糖基化N HRSV蛋白在SNX2负载的聚集体中检测到，细胞内细丝从它们的外表面突出，并且VPS26，另一种逆转录组分，也在包涵体和顺应性形状的结构中发现。与SNX2相似，TGN 46也与HRSV M和N蛋白共定位于质膜上的丝状结构中。细胞分级分离显示SNX2在含有HRSV M和N蛋白的级分中富集。SNX 1和2的沉默与病毒蛋白质、HRSV包含体大小、合胞体形成和后代产生的减少有关。结果表明，HRSV结构蛋白M和N处于分泌途径，SNX2在HRSV结构蛋白向组装位点的运输中起重要作用。
The present study contributes new knowledge to understand HRSV assembly by providing evidence that nonglycosylated structural proteins M and N interact with elements of the secretory pathway, shedding light on their intracellular traffic. To the best of our knowledge, the present contribution is important given the scarcity of studies about the traffic of HRSV nonglycosylated proteins, especially by pointing to the involvement of SNX2, a retromer component, in the HRSV assembly process. Human respiratory syncytial virus (HRSV) envelope glycoproteins traffic to assembly sites through the secretory pathway, while nonglycosylated proteins M and N are present in HRSV inclusion bodies but must reach the plasma membrane, where HRSV assembly happens. Little is known about how nonglycosylated HRSV proteins reach assembly sites. Here, we show that HRSV M and N proteins partially colocalize with the Golgi marker giantin, and the glycosylated F and nonglycosylated N proteins are closely located in the trans-Golgi, suggesting their interaction in that compartment. Brefeldin A compromised the trafficking of HRSV F and N proteins and inclusion body sizes, indicating that the Golgi is important for both glycosylated and nonglycosylated HRSV protein traffic. HRSV N and M proteins colocalized and interacted with sorting nexin 2 (SNX2), a retromer component that shapes endosomes in tubular structures. Glycosylated F and nonglycosylated N HRSV proteins are detected in SNX2-laden aggregates with intracellular filaments projecting from their outer surfaces, and VPS26, another retromer component, was also found in inclusion bodies and filament-shaped structures. Similar to SNX2, TGN46 also colocalized with HRSV M and N proteins in filamentous structures at the plasma membrane. Cell fractionation showed enrichment of SNX2 in fractions containing HRSV M and N proteins. Silencing of SNX1 and 2 was associated with reduction in viral proteins, HRSV inclusion body size, syncytium formation, and progeny production. The results indicate that HRSV structural proteins M and N are in the secretory pathway, and SNX2 plays an important role in the traffic of HRSV structural proteins toward assembly sites.