猪瘟病毒衣壳蛋白与宿主丝/苏氨酸蛋白激酶N1相互作用的鉴定

Identification of the interaction between classical swine fever virus C protein and porcine PKN1

为筛选并鉴定与猪瘟病毒(CSFV)衣壳(C)蛋白相互作用的宿主蛋白,本研究采用酵母双杂交技术以CSFV C蛋白为诱饵从猪外周血单个核细胞(PBMC)c DNA表达文库中筛选与之相互作用的宿主蛋白,共筛选到6种蛋白,分别为ATP5B、SON3、PKN1、PCBP1、RPS20和IQGAP1,根据Gene Ontology分析结果,这些蛋白分别参与细胞的增殖和代谢等过程。本研究选取丝/苏氨酸蛋白激酶N1(PKN1)进行进一步验证,经酵母共转化试验、免疫共沉淀试验和GST pull-down试验证实,宿主PKN1蛋白与CSFV C蛋白之间存在特异性结合。本研究首次证明PKN1与CSFV蛋白之间的相互作用关系,为进一步研究PKN1蛋白在CSFV感染过程中发挥的作用奠定了基础。

To identify the porcine proteins that interacting with the classical swine fever virus（CSFV） capsid（C） protein, a total of 6 potential interacting partners were scanned with the C protein as bait protein from porcine peripheral blood mononuclear cell（PBMC） expression library by yeast two-hybrid system（Y2H）. The Gene-Ontology（GO） analysis showed that the proteins were found to be involved in cellular process, metabolic process, and localization, respectively. In addition, the interaction of serine/threonine-protein kinase N1（PKN1） protein and C protein was further verified and confirmed by co-transformation, GST pull-down and co-immunoprecipitation assays. These results demonstrated that PKN1 was one of the proteins that interacted with CSFV C protein, which would facilitate future study of the role of PKN1 in CSFV replication.