Rho GTP酶和肌动蛋白细胞骨架对新城疫病毒在细胞间传播的影响

Effects of Rho GTPase and actin cytoskeleton on the intercellular spread of Newcastle disease virus

为探究Rho GTP酶及肌动蛋白细胞骨架对新城疫病毒(NDV)诱导的细胞间膜融合以及病毒复制和扩散的影响,本研究利用特异性针对Rho GTP酶和肌动蛋白细胞骨架的抑制剂处理细胞,评估其对NDV复制以及细胞间膜融合的影响,结果显示,抑制剂处理能够显著抑制NDV在细胞中的复制滴度以及病毒引起的细胞间膜融合。激光共聚焦观察结果显示,NDV的融合蛋白(F)与细胞的肌动蛋白存在共定位分布。通过反向遗传技术构建了表达绿色荧光蛋白的重组NDV,并通过共感染培养试验证实了Rho GTP酶和肌动蛋白细胞骨架参与了NDV在细胞间的扩散和传播。本研究结果为深入探索Rho GTP酶和肌动蛋白细胞骨架在NDV感染生活周期中的作用奠定了基础。

In this study,in order to explore the effects of Rho GTPase and actin cytoskeleton on NDV-induced intercellular membrane fusion and viral replication and spread,specific inhibitors target to Rho GTPase and actin cytoskeleton were firstly used to treat cells and evaluated their effects on replication of NDV and intercellular membrane fusion induced by NDV.The results showed that inhibitor treatment could significantly inhibit the replication titer of NDv in cells and the intercellular membrane fusion induced by this virus.Furthermore,the observation results of laser scanning confocal microscope showed that the fusion protein(F)of NDV and the actin of the cells were co-localized.Finally,the recombinant NDV expressing green fluorescent protein was rescued by reverse genetic technique,and the co-infection culture experiment revealed that Rho GTPase and actin cytoskeleton were involved in the transmission and spread of NDV among cells.The results of this study provided information for deep understanding of the role of Rho GTPase and actin cytoskeleton in the life cycle of NDV infection.