A型流感病毒PB1-F2蛋白与MOAP-1的相互作用

【目的】探讨A型流感病毒PB1-F2蛋白和人类凋亡调节因子1(MOAP-1)之间的相互作用。【方法】构建pACT2-MOAP-1重组质粒,与pGBKT7-PB1-F2质粒共转化酵母AH109,检测转化菌在四缺培养基的生长情况及β半乳糖苷酶报告基因的活性;利用GST pull-down和免疫共沉淀(Co-IP)技术进一步验证PB1-F2与宿主细胞蛋白MOAP-1的相互作用;通过过表达PB1-F2和MOAP-1,检测PB1-F2对MOAP-1蛋白表达水平的影响。【结果】酵母双杂交结果表明,PB1-F2和MOAP-1可以在酵母细胞内特异性结合。GST pull-down和Co-IP实验也进一步证实了这两种蛋白的相互作用,而且PB1-F2可上调外源MOAP-1的蛋白水平。【结论】流感病毒PB1-F2与MOAP-1存在相互作用,PB1-F2可能通过与MOAP-1的相互作用参与调控细胞生长及凋亡过程。

PB1-F2 protein of influenza A virus interacts with human MOAP-1 protein

[Objective] To identify the interaction between influenza A virus PB1-F2 and human modulator of apoptosis 1(MOAP-1).[Methods] The recombinant plasmid pACT2-MOAP-1 was constructed and co-transformed into yeast AHl09 with pGBKT7-PB1-F2.The growth of the co-transformants on quadruple dropout medium and β-galactosidase activity of the reporter gene were tested.We further confirmed the interaction of cellular protein MOAP-1 and PB1-F2 by glutathione S-transferase(GST)pull-down and co-immunoprecipitation(Co-IP) assays.In addition,we investigated the effect of PB1-F2 on MOAP-1 protein level by Western blot.[Results] The results of yeast two-hybrid assay showed that MOAP-1 specifically interacted with PB1-F2 in yeast cells.Furthermore,the binding of MOAP-1 with PB1-F2 was demonstrated by glutathione S-transferase pull-down and Co-IP assays.PB1-F2 could upregulate exogenous MOAP-1 protein level.[Conclusion] These results suggested that influenza virus PB1-F2 interacted with MOAP-1 and it might be involved in the regulation of cell growth and apoptosis via association with MOAP-1.