Department of Medicinal Chemistry and Molecular Pharmacology, Purdue University, West Lafayette, IN, 47907, USA; Department of Molecular Pharmacology, Purdue University, West Lafayette, IN, 47907, USA; Purdue Institute of Inflammation, Immunology and Infectious Disease, Purdue University, West Lafayette, IN, 47907, USA.
Department of Biological Sciences, Purdue University, West Lafayette, IN, 47907, USA; Purdue Institute of Inflammation, Immunology and Infectious Disease, Purdue University, West Lafayette, IN, 47907, USA.
Virology. 2020 Feb;541:52-62. doi: 10.1016/j.virol.2019.11.018. Epub 2019 Dec 6.
Zika virus (ZIKV) nonstructural protein 5 (NS5) plays a critical role in viral RNA replication and mediates key virus-host cell interactions. As with other flavivirus NS5 proteins, ZIKV NS5 is primarily found in the nucleus. We previously reported that the NS5 protein of dengue virus, another flavivirus, localized to centrosomes during cell division. Here we show that ZIKV NS5 also relocalizes from the nucleus to centrosomes during mitosis. In infected cells with supernumerary centrosomes, NS5 was present at all centrosomes. Transient expression of NS5 in uninfected cells confirmed that centrosomal localization was independent of other viral proteins. Live-cell imaging demonstrated that NS5-GFP accumulated at centrosomes shortly after break down of nuclear membrane and remained there through mitosis. Cells expressing NS5-GFP took longer to complete mitosis than control cells. Finally, an analysis of ZIKV NS5 binding partners revealed several centrosomal proteins, providing potential direct links between NS5 and centrosomes.
寨卡病毒（ZIKV）非结构蛋白 5（NS5）在病毒 RNA 复制中发挥关键作用，并介导关键的病毒-宿主细胞相互作用。与其他黄病毒 NS5 蛋白一样，ZIKV NS5 主要存在于细胞核中。我们之前报道过，另一种黄病毒登革热病毒的 NS5 蛋白在细胞分裂过程中定位于中心体。在这里，我们表明 ZIKV NS5 在有丝分裂过程中也从细胞核重新定位到中心体。在具有多余中心体的感染细胞中，NS5 存在于所有中心体上。未感染细胞中 NS5 的瞬时表达证实了中心体定位不依赖于其他病毒蛋白。活细胞成像表明，NS5-GFP 在核膜破裂后不久就聚集在中心体上，并在有丝分裂过程中一直存在于此。表达 NS5-GFP 的细胞比对照细胞完成有丝分裂所需的时间更长。最后，对 ZIKV NS5 结合伙伴的分析揭示了几种中心体蛋白，为 NS5 和中心体之间提供了潜在的直接联系。
