Pirinçal Ayşegül, Turan Kadir
Marmara University, Institute of Health Sciences, Istanbul, Turkey.
Marmara University, Faculty of Pharmacy, Department of Basic Pharmaceutical Sciences, Istanbul, Turkey.
Genet Mol Biol. 2021 Mar 22;44(1):e20200158. doi: 10.1590/1678-4685-GMB-2020-0158. eCollection 2021.
Influenza A viruses (IAV) are enveloped viruses carrying a single-stranded negative-sense RNA genome. Detection of host proteins having a relationship with IAV and revealing of the role of these proteins in the viral replication are of great importance in keeping IAV infections under control. Consequently, the importance of human DDX56, which is determined to be associated with a viral NS1 with a yeast two-hybrid assay, was investigated for IAV replication. The viral replication in knocked down cells for the DDX56 gene was evaluated. The NS1 was co-precipitated with the DDX56 protein in lysates of cells transiently expressing DDX56 and NS1 or infected with the viruses, showing that NS1 and DDX56 interact in mammalian cells. Viral NS1 showed a tendency to co-localize with DDX56 in the cells, transiently expressing both of these proteins, which supports the IP and two-hybrid assays results. The data obtained with in silico predictions supported the in vitro protein interaction results. The viral replication was significantly reduced in the DDX56-knockdown cells comparing with that in the control cells. In conclusion, human DDX56 protein interacts with the IAV NS1 protein in both yeast and mammalian cells and has a positive regulatory effect on IAV replication. However, the mechanism of DDX56 on IAV replication requires further elucidation.
甲型流感病毒(IAV)是包膜病毒,携带单链负义RNA基因组。检测与IAV相关的宿主蛋白并揭示这些蛋白在病毒复制中的作用对于控制IAV感染至关重要。因此,研究了通过酵母双杂交试验确定与病毒NS1相关的人类DDX56在IAV复制中的重要性。评估了DDX56基因敲低细胞中的病毒复制情况。在瞬时表达DDX56和NS1的细胞裂解物或感染病毒的细胞裂解物中,NS1与DDX56蛋白共沉淀,表明NS1和DDX56在哺乳动物细胞中相互作用。在瞬时表达这两种蛋白的细胞中,病毒NS1显示出与DDX56共定位的趋势,这支持了免疫沉淀和双杂交试验结果。计算机预测获得的数据支持了体外蛋白质相互作用结果。与对照细胞相比,DDX56敲低细胞中的病毒复制显著减少。总之,人类DDX56蛋白在酵母和哺乳动物细胞中均与IAV NSI蛋白相互作用,并对IAV复制具有正向调节作用。然而,DDX56对IAV复制的机制需要进一步阐明。