Instituto de Biotecnología, CICVyA, Instituto Nacional de Tecnología Agropecuaria (IB-INTA), Las cabañas y Los Reseros s/n. Hurlingham Cp 1686, Buenos Aires, Argentina.
Virology. 2012 Sep 1;430(2):81-9. doi: 10.1016/j.virol.2012.04.016. Epub 2012 May 18.
The in vivo subcellular localization of Mal de Río Cuarto virus (MRCV, Fijivirus, Reoviridae) non-structural proteins fused to GFP was analyzed by confocal microscopy. P5-1 showed a cytoplasmic vesicular-like distribution that was lost upon deleting its PDZ binding TKF motif, suggesting that P5-1 interacts with cellular PDZ proteins. P5-2 located at the nucleus and its nuclear import was affected by the deletion of its basic C-termini. P7-1 and P7-2 also entered the nucleus and therefore, along with P5-2, could function as regulators of host gene expression. P6 located in the cytoplasm and in perinuclear cloud-like inclusions, was driven to P9-1 viroplasm-like structures and co-localized with P7-2, P10 and α-tubulin, suggesting its involvement in viroplasm formation and viral intracellular movement. Finally, P9-2 was N-glycosylated and located at the plasma membrane in association with filopodia-like protrusions containing actin, suggesting a possible role in virus cell-to-cell movement and spread.
通过共聚焦显微镜分析了融合 GFP 的马尔德里奥库图病毒（MRCV、Fijivirus、正粘病毒科）非结构蛋白的体内亚细胞定位。P5-1 表现出细胞质囊泡样分布，删除其 PDZ 结合 TKF 基序后这种分布消失，表明 P5-1 与细胞 PDZ 蛋白相互作用。P5-2 位于细胞核内，其核输入受其碱性 C 末端缺失的影响。P7-1 和 P7-2 也进入细胞核，因此与 P5-2 一起可以作为宿主基因表达的调节剂。P6 位于细胞质和核周云状包涵体中，被驱动到 P9-1 类病毒体样结构中，并与 P7-2、P10 和 α-微管蛋白共定位，表明其参与类病毒体形成和病毒细胞内运动。最后，P9-2 发生 N-糖基化并与含有肌动蛋白的丝状伪足相关联位于质膜上，提示其可能在病毒细胞间运动和传播中发挥作用。
