Department of Biopharmacy, College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou 310018, China.
State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, China.
Viruses. 2022 Jun 21;14(7):1347. doi: 10.3390/v14071347.
Increasing evidence suggests that the polymerase acidic (PA) protein of influenza A viruses plays an important role in viral replication and pathogenicity. However, information regarding the interaction(s) of host factors with PA is scarce. By using a yeast two-hybrid screen, we identified a novel host factor, aryl hydrocarbon receptor nuclear translocator (ARNT), that interacts with the PA protein of the H5N1 virus. The interaction between PA and human ARNT was confirmed by co-immunoprecipitation and immunofluorescence microscopy. Moreover, overexpression of ARNT downregulated the polymerase activity and inhibited virus propagation, whereas knockdown of ARNT significantly increased the polymerase activity and virus replication. Mechanistically, overexpression of ARNT resulted in the accumulation of PA protein in the nucleus and inhibited both the replication and transcription of the viral genome. Interaction domain mapping revealed that the bHLH/PAS domain of ARNT mainly interacted with the C-terminal domain of PA. Together, our results demonstrate that ARNT inhibits the replication of the H5N1 virus and could be a target for the development of therapeutic strategies against H5N1 influenza viruses.
越来越多的证据表明，甲型流感病毒的聚合酶酸性（PA）蛋白在病毒复制和致病性方面发挥着重要作用。然而，关于宿主因子与 PA 的相互作用的信息却很少。通过使用酵母双杂交筛选，我们鉴定出一种新型宿主因子，芳香烃受体核转位蛋白（ARNT），它与 H5N1 病毒的 PA 蛋白相互作用。PA 和人 ARNT 之间的相互作用通过共免疫沉淀和免疫荧光显微镜得到了证实。此外，ARNT 的过表达下调了聚合酶活性并抑制了病毒的繁殖，而 ARNT 的敲低则显著增加了聚合酶活性和病毒复制。从机制上讲，ARNT 的过表达导致 PA 蛋白在核内积累，并抑制病毒基因组的复制和转录。相互作用域作图显示，ARNT 的 bHLH/PAS 结构域主要与 PA 的 C 末端结构域相互作用。总之，我们的研究结果表明，ARNT 抑制了 H5N1 病毒的复制，可能成为针对 H5N1 流感病毒的治疗策略的靶点。
