猪圆环病毒2型Cap蛋白互作宿主核蛋白的筛选与鉴定

Screening and identification of porcine circovirus type 2 Cap protein-interacting host nuclear proteins

为探究在猪圆环病毒2型(PCV2)入核复制过程中发挥生物学功能的宿主核相关蛋白,本研究借助免疫沉淀技术联合蛋白质谱鉴定技术,从宿主细胞核膜提取物中筛选到了可与PCV2 Cap蛋白发生潜在互作的2个蛋白质:高迁移率族蛋白1(HMGB1)和精氨酸酶1(ARG1)。通过进一步的免疫共沉淀试验及激光共聚焦试验,证明了HMGB1、ARG1与PCV2 Cap蛋白的相互作用,且互作与Cap蛋白的核定位信号(NLS)无关。该研究为进一步探索PCV2入核复制的机制解析提供了重要信息。

To investigate the host nuclear-associated proteins that play a biological function in the nuclear entry of porcine circovirus type 2(PCV2),a coimmunoprecipitation combined with liquid chromatography mass spectrometry approach was used to screen two proteins from host cell nuclear membrane extracts for potential interactions with PCV2 Cap proteins,the high mobility group protein 1(HMGB1)and arginase 1(ARG1),respectively.The interaction of HMGB1 and ARG1 with PCV2 Cap proteins was further demonstrated by coimmunoprecipitation assays and laser confocal assays,furthermore,the interactions were independent of the nuclear localization signal(NLS)of Cap proteins.This study provides important information for further exploring the mechanism of PCV2 nuclear entry.