MRC Laboratory for Molecular Cell Biology, University College London, London, UK.
Institute of Microbiology and Infection, University of Birmingham, Birmingham, UK.
Cell Microbiol. 2021 Aug;23(8):e13334. doi: 10.1111/cmi.13334. Epub 2021 May 3.
The replication and assembly of vaccinia virus (VACV), the prototypic poxvirus, occurs exclusively in the cytoplasm of host cells. While the role of cellular cytoskeletal components in these processes remains poorly understood, vimentin-a type III intermediate filament-has been shown to associate with viral replication sites and to be incorporated into mature VACV virions. Here, we employed chemical and genetic approaches to further investigate the role of vimentin during the VACV lifecycle. The collapse of vimentin filaments, using acrylamide, was found to inhibit VACV infection at the level of genome replication, intermediate- and late-gene expression. However, we found that CRISPR-mediated knockout of vimentin did not impact VACV replication. Combining these tools, we demonstrate that acrylamide treatment results in the formation of anti-viral granules (AVGs) known to mediate translational inhibition of many viruses. We conclude that vimentin is dispensable for poxvirus replication and assembly and that acrylamide, as a potent inducer of AVGs during VACV infection, serves to bolster cell's anti-viral response to poxvirus infection.
痘苗病毒（VACV）是典型的正痘病毒，其复制和组装仅发生在宿主细胞的细胞质中。虽然细胞骨架成分在这些过程中的作用仍知之甚少，但波形蛋白 - 一种 III 型中间丝 - 已被证明与病毒复制位点结合，并被纳入成熟的 VACV 病毒粒子中。在这里，我们采用化学和遗传方法进一步研究了波形蛋白在 VACV 生命周期中的作用。使用丙烯酰胺使波形蛋白丝塌陷，发现会抑制基因组复制、中期和晚期基因表达水平的 VACV 感染。然而，我们发现 CRISPR 介导的波形蛋白敲除并不影响 VACV 复制。结合这些工具，我们证明丙烯酰胺处理会导致抗病毒颗粒（AVGs）的形成，已知 AVGs 可抑制许多病毒的翻译。我们得出结论，波形蛋白对于痘病毒的复制和组装是可有可无的，而丙烯酰胺作为 VACV 感染期间抗病毒颗粒的有效诱导剂，可增强细胞对痘病毒感染的抗病毒反应。
