African Swine Fever Vaccinology Group, The Pirbright Institute, Ash Road, Pirbright, Woking, Surrey GU24 0NF, UK.
Bioimaging, The Pirbright Institute, Ash Road, Pirbright, Woking, Surrey GU24 0NF, UK.
Viruses. 2021 Jan 8;13(1):77. doi: 10.3390/v13010077.
African swine fever virus (ASFV) is a highly contagious pathogen which causes a lethal haemorrhagic fever in domestic pigs and wild boar. The large, double-stranded DNA virus replicates in perinuclear cytoplasmic replication sites known as viral factories. These factories are complex, multi-dimensional structures. Here we investigated the protein and membrane compartments of the factory using super-resolution and electron tomography. Click IT chemistry in combination with stimulated emission depletion (STED) microscopy revealed a reticular network of newly synthesized viral proteins, including the structural proteins p54 and p34, previously seen as a pleomorphic ribbon by confocal microscopy. Electron microscopy and tomography confirmed that this network is an accumulation of membrane assembly intermediates which take several forms. At early time points in the factory formation, these intermediates present as small, individual membrane fragments which appear to grow and link together, in a continuous progression towards new, icosahedral virions. It remains unknown how these membranes form and how they traffic to the factory during virus morphogenesis.
非洲猪瘟病毒（ASFV）是一种高度传染性的病原体，可导致家猪和野猪发生致命性出血热。这种大型双链 DNA 病毒在称为病毒工厂的核周细胞质复制位点中复制。这些工厂是复杂的、多维的结构。在这里，我们使用超分辨率和电子断层扫描技术研究了工厂的蛋白质和膜区室。点击 IT 化学与受激发射耗散（STED）显微镜相结合，揭示了新合成的病毒蛋白的网状网络，包括结构蛋白 p54 和 p34，之前通过共聚焦显微镜观察到的是一种多形性的带状结构。电子显微镜和断层扫描证实，该网络是膜组装中间体的积累，其呈现出多种形式。在工厂形成的早期阶段，这些中间体表现为小的、单个的膜片段，似乎在不断生长并连接在一起，朝着新的二十面体病毒颗粒前进。目前尚不清楚这些膜是如何形成的，以及它们在病毒形态发生过程中是如何运输到工厂的。
