MRC-University of Glasgow Centre for Virus Research, Glasgow G61 1QH, UK.
Center for Autoimmune Diseases Research-CREA, School of Medicine and Health Sciences, Universidad del Rosario, 110010 Bogotá, Colombia.
Viruses. 2020 May 9;12(5):524. doi: 10.3390/v12050524.
Zika virus (ZIKV; Flaviviridae) is a mosquito-borne flavivirus shown to cause fetal abnormalities collectively known as congenital Zika syndrome and Guillain-Barré syndrome in recent outbreaks. Currently, there is no specific treatment or vaccine available, and more effort is needed to identify cellular factors in the viral life cycle. Here, we investigated interactors of ZIKV envelope (E) protein by combining protein pull-down with mass spectrometry. We found that E interacts with the endoplasmic reticulum (ER) resident chaperone, glucose regulated protein 78 (GRP78). Although other flaviviruses are known to co-opt ER resident proteins, including GRP78, to enhance viral infectivity, the role ER proteins play during the ZIKV life cycle is yet to be elucidated. We showed that GRP78 levels increased during ZIKV infection and localised to sites coincident with ZIKV E staining. Depletion of GRP78 using specific siRNAs significantly reduced reporter-virus luciferase readings, viral protein synthesis, and viral titres. Additionally, GRP78 depletion reduced the ability of ZIKV to disrupt host cell translation and altered the localisation of viral replication factories, though there was no effect on viral RNA synthesis. In summary, we showed GRP78 is a vital host-factor during ZIKV infection, which may be involved in the coordination of viral replication factories.
Zika 病毒(ZIKV;黄病毒科)是一种通过蚊子传播的黄病毒,最近的疫情爆发表明其会导致胎儿畸形,统称为先天性 Zika 综合征和格林-巴利综合征。目前,尚无特定的治疗方法或疫苗,需要更多的努力来确定病毒生命周期中的细胞因子。在这里,我们通过将蛋白质下拉与质谱相结合,研究了 ZIKV 包膜(E)蛋白的相互作用蛋白。我们发现 E 与内质网(ER)驻留伴侣葡萄糖调节蛋白 78(GRP78)相互作用。虽然其他黄病毒也已知会劫持 ER 驻留蛋白,包括 GRP78,以增强病毒感染力,但 ER 蛋白在 ZIKV 生命周期中的作用尚未阐明。我们表明,在 ZIKV 感染过程中,GRP78 的水平增加,并定位于与 ZIKV E 染色重合的部位。使用特异性 siRNA 耗尽 GRP78 会显著降低报告病毒荧光素酶读数、病毒蛋白合成和病毒滴度。此外,GRP78 耗尽会降低 ZIKV 破坏宿主细胞翻译的能力,并改变病毒复制工厂的定位,尽管对病毒 RNA 合成没有影响。总之,我们表明 GRP78 是 ZIKV 感染过程中的重要宿主因子,可能参与病毒复制工厂的协调。