de Wilde Adriaan H, Zevenhoven-Dobbe Jessika C, Beugeling Corrine, Chatterji Udayan, de Jong Danielle, Gallay Philippe, Szuhai Karoly, Posthuma Clara C, Snijder Eric J
Molecular Virology Laboratory, Department of Medical Microbiology, Leiden University Medical Center, Leiden, The Netherlands.
Molecular Virology Laboratory, Department of Medical Microbiology, Leiden University Medical Center, Leiden, The Netherlands.
Virology. 2018 Apr;517:148-156. doi: 10.1016/j.virol.2017.11.022. Epub 2017 Dec 15.
Cyclophilin A (CypA) is an important host factor in the replication of a variety of RNA viruses. Also the replication of several nidoviruses was reported to depend on CypA, although possibly not to the same extent. These prior studies are difficult to compare, since different nidoviruses, cell lines and experimental set-ups were used. Here, we investigated the CypA dependence of three distantly related nidoviruses that can all replicate in Huh7 cells: the arterivirus equine arteritis virus (EAV), the alphacoronavirus human coronavirus 229E (HCoV-229E), and the betacoronavirus Middle East respiratory syndrome coronavirus (MERS-CoV). The replication of these viruses was compared in the same parental Huh7 cells and in CypA-knockout Huh7 cells generated using CRISPR/Cas9-technology. CypA depletion reduced EAV yields by ~ 3-log, whereas MERS-CoV progeny titers were modestly reduced (3-fold) and HCoV-229E replication was unchanged. This study reveals that the replication of nidoviruses can differ strikingly in its dependence on cellular CypA.
亲环蛋白A（CypA）是多种RNA病毒复制过程中的一个重要宿主因子。据报道，几种巢病毒的复制也依赖于CypA，尽管依赖程度可能不尽相同。由于使用了不同的巢病毒、细胞系和实验设置，这些先前的研究难以进行比较。在此，我们研究了三种远缘相关的巢病毒在CypA方面的依赖性，这三种病毒均可在Huh7细胞中复制：动脉炎病毒马动脉炎病毒（EAV）、甲型冠状病毒人冠状病毒229E（HCoV-229E）以及乙型冠状病毒中东呼吸综合征冠状病毒（MERS-CoV）。在相同的亲本Huh7细胞以及使用CRISPR/Cas9技术构建的CypA基因敲除Huh7细胞中，对这些病毒的复制情况进行了比较。CypA缺失使EAV产量降低了约3个对数级，而MERS-CoV子代滴度略有降低（3倍），HCoV-229E的复制则没有变化。这项研究表明，巢病毒的复制对细胞CypA的依赖性可能存在显著差异。
