Das Provas, Saha Shekhar, Chandra Sunandini, Das Alakesh, Dey Sumit K, Das Mahua R, Sen Shamik, Sarkar Debi P, Jana Siddhartha S
Department of Biological Chemistry, Indian Association for the Cultivation of Science, Jadavpur, Kolkata-700032.
Department of Biochemistry, University of Delhi South Campus, New Delhi-1100021.
Sci Rep. 2015 May 20;5:10395. doi: 10.1038/srep10395.
Enveloped viruses enter host cells through membrane fusion and the cells in turn alter their shape to accommodate components of the virus. However, the role of nonmuscle myosin II of the actomyosin complex of host cells in membrane fusion is yet to be understood. Herein, we show that both (-) blebbistatin, a specific inhibitor of nonmuscle myosin II (NMII) and small interfering RNA markedly augment fusion of Sendai virus (SeV), with chinese hamster ovary cells and human hepatocarcinoma cells. Inhibition of RLC phosphorylation using inhibitors against ROCK, but not PKC and MRCK, or overexpression of phospho-dead mutant of RLC enhances membrane fusion. SeV infection increases cellular stiffness and myosin light chain phosphorylation at two hour post infection. Taken together, the present investigation strongly indicates that Rho-ROCK-NMII contractility signaling pathway may provide a physical barrier to host cells against viral fusion.
包膜病毒通过膜融合进入宿主细胞，宿主细胞会相应地改变其形状以容纳病毒成分。然而，宿主细胞肌动球蛋白复合体中的非肌肉肌球蛋白II在膜融合中的作用尚不清楚。在此，我们发现，非肌肉肌球蛋白II（NMII）的特异性抑制剂（-）blebbistatin和小干扰RNA均可显著增强仙台病毒（SeV）与中国仓鼠卵巢细胞和人肝癌细胞的融合。使用针对ROCK而非PKC和MRCK的抑制剂抑制RLC磷酸化，或过表达RLC的磷酸化缺失突变体均可增强膜融合。SeV感染在感染后两小时增加细胞硬度和肌球蛋白轻链磷酸化。综上所述，本研究强烈表明，Rho-ROCK-NMII收缩信号通路可能为宿主细胞抵御病毒融合提供物理屏障。
