College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, 712100, China.
College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, 712100, China.
Vet Microbiol. 2021 Apr;255:109034. doi: 10.1016/j.vetmic.2021.109034. Epub 2021 Mar 5.
Classical swine fever virus (CSFV), an enveloped virus belonging to the genus Pestivirus of the Flaviviridae family, utilizes cell host factors for its own replication. ARFGAP1, GTPase activating protein of ADP-ribosylation factor 1, regulates COP I vesicle formation and function in cells and is involved in the life cycle of several viruses. However, the effect of ARFGAP1 on the infection of CSFV has not been illustrated. Here we showed that inhibition of ARFGAP1 either by QS11 or by lentivirus-mediated silencing repressed CSFV replication. While, subsequent experiments revealed that CSFV production were increased in cells with sufficient ARFGAP1 expression. However, ARFGAP1 was not involved in CSFV binding, entry, access to cell vesicles, and RNA replication during the early stages of infection. Then, we showed that ARFGAP1 interacted with the viral protein of NS5A, measured by immunoprecipitation, GST-pulldown, and confocal microscopy assays. Furthermore, we revealed that ARFGAP1 could alleviated CSFV NS5A-induced endoplasmic reticulum stress (ERS). Altogether, these results demonstrate that ARFGAP1, a NS5A binding protein, is involved in CSFV replication.
经典猪瘟病毒（CSFV）是一种囊膜病毒，属于黄病毒科瘟病毒属，利用细胞宿主因子进行自身复制。ARFGAP1 是 ADP-核糖基化因子 1 的 GTP 酶激活蛋白，调节细胞中 COP I 囊泡的形成和功能，并参与几种病毒的生命周期。然而，ARFGAP1 对 CSFV 感染的影响尚未阐明。在这里，我们表明通过 QS11 或慢病毒介导的沉默抑制 ARFGAP1 抑制了 CSFV 的复制。然而，随后的实验表明，在 ARFGAP1 表达充足的细胞中，CSFV 的产量增加。然而，ARFGAP1 不参与 CSFV 结合、进入、进入细胞囊泡和感染早期的 RNA 复制。然后，我们通过免疫沉淀、GST 下拉和共聚焦显微镜检测证实了 ARFGAP1 与病毒蛋白 NS5A 的相互作用。此外，我们还揭示了 ARFGAP1 可以减轻 CSFV NS5A 诱导的内质网应激（ERS）。总之，这些结果表明，作为 NS5A 结合蛋白的 ARFGAP1 参与了 CSFV 的复制。
