Instituto de Virología "Dr. JM Vanella", Facultad de Ciencias Médicas, Universidad Nacional de Córdoba, Córdoba, Argentina.
Centro de Investigaciones en Ciencias de la Tierra CICTERRA, CONICET, Universidad Nacional de Córdoba, Córdoba, Argentina.
Sci Rep. 2017 Jul 18;7(1):5757. doi: 10.1038/s41598-017-05983-w.
Pixuna virus (PIXV) is an enzootic member of the Venezuelan Equine Encephalitis Virus complex and belongs to the New World cluster of alphaviruses. Herein we explore the role of the cellular cytoskeleton during PIXV replication. We first identified that PIXV undergoes an eclipse phase consisting of 4 h followed by 20 h of an exponential phase in Vero cells. The infected cells showed morphological changes due to structural modifications in actin microfilaments (MFs) and microtubules (MTs). Cytoskeleton-binding agents, that alter the architecture and dynamics of MFs and MTs, were used to study the role of cytoskeleton on PIXV replication. The virus production was significantly affected (p < 0.05) after treatment with paclitaxel or nocodazole due to changes in the MTs network. Interestingly, disassembly of MFs with cytochalasin D, at early stage of PIXV replication cycle, significantly increased the virus yields in the extracellular medium (p < 0.005). Furthermore, the stabilization of actin network with jasplakinolide had no effect on virus yields. Our results demonstrate that PIXV relies not only on intact MTs for the efficient production of virus, but also on a dynamic actin network during the early steps of viral replication.
皮库纳病毒（PIXV）是委内瑞拉马脑炎病毒复合体中的内栖成员，属于新世界组的甲病毒属。在此，我们探讨了细胞骨架在 PIXV 复制过程中的作用。我们首先确定 PIXV 在 Vero 细胞中经历了一个包含 4 小时潜伏期和 20 小时指数期的潜伏期。受感染的细胞由于肌动蛋白微丝（MFs）和微管（MTs）的结构修饰而发生形态变化。使用细胞骨架结合剂来改变 MFs 和 MTs 的结构和动力学，以研究细胞骨架对 PIXV 复制的作用。紫杉醇或诺考达唑处理后，病毒产量显著受到影响（p<0.05），这是由于 MTs 网络发生变化所致。有趣的是，在 PIXV 复制周期的早期用细胞松弛素 D 分解 MFs，会显著增加细胞外介质中的病毒产量（p<0.005）。此外，用 Jasplakinolide 稳定肌动蛋白网络对病毒产量没有影响。我们的结果表明，PIXV 不仅依赖于完整的 MTs 来高效地产生病毒，而且在病毒复制的早期阶段还依赖于动态的肌动蛋白网络。
