Emory Vaccine Center, Yerkes National Primate Research Center, 954 Gatewood Road NE, Atlanta, GA 30329, USA.
Department of Biochemistry and Microbiology, Institute of Chemical Technology, Technicka 3, 166 28 Prague, Czech Republic.
Virology. 2014 Jan 20;449:109-19. doi: 10.1016/j.virol.2013.11.006. Epub 2013 Nov 28.
The intracellular transport of Mason-Pfizer monkey virus (M-PMV) assembled capsids from the pericentriolar region to the plasma membrane (PM) requires trafficking of envelope glycoprotein (Env) to the assembly site via the recycling endosome. However, it is unclear if Env-containing vesicles play a direct role in trafficking capsids to the PM. Using live cell microscopy, we demonstrate, for the first time, anterograde co-transport of Gag and Env. Nocodazole disruption of microtubules had differential effects on Gag and Env trafficking, with pulse-chase assays showing a delayed release of Env-deficient virions. Particle tracking demonstrated an initial loss of linear movement of GFP-tagged capsids and mCherry-tagged Env, followed by renewed movement of Gag but not Env at 4h post-treatment. Thus, while delayed capsid trafficking can occur in the absence of microtubules, efficient anterograde transport of capsids appears to be mediated by microtubule-associated Env-containing vesicles.
从中心体周围区域到质膜（PM）的 Mason-Pfizer 猴病毒（M-PMV）组装衣壳的细胞内运输需要包膜糖蛋白（Env）通过再循环内体转运到组装部位。然而，Env 包含的囊泡是否直接参与将衣壳运输到 PM 尚不清楚。使用活细胞显微镜，我们首次证明了 Gag 和 Env 的顺行共转运。微管的诺考达唑破坏对 Gag 和 Env 运输有不同的影响，脉冲追踪试验显示 Env 缺陷型病毒粒子的释放延迟。粒子追踪显示 GFP 标记的衣壳和 mCherry 标记的 Env 的线性运动最初丢失，随后在处理后 4 小时 Gag 重新运动，但 Env 没有运动。因此，虽然在没有微管的情况下可以发生衣壳运输延迟，但衣壳的有效顺行运输似乎是由微管相关的含有 Env 的囊泡介导的。
