Plum Island Animal Disease Center, ARS, USDA, Greenport, NY 11944, USA.
Oak Ridge Institute for Science and Education (ORISE), Oak Ridge, TN 37830, USA.
Viruses. 2019 Mar 29;11(4):307. doi: 10.3390/v11040307.
Classical swine fever virus (CSFV) E2 protein, the major virus structural glycoprotein, is an essential component of the viral envelope. E2 is involved in virus absorption, induction of a protective immune response and is critical for virulence in swine. Using the yeast two-hybrid system, we identified protein phosphatase 1 catalytic subunit beta (PPP1CB), which is part of the Protein Phosphatase 1 (PP1) complex, as a specific binding host partner for E2. We further confirmed the occurrence of this interaction in CSFV-infected swine cells by using two independent methodologies: Co-immunoprecipitation and Proximity Ligation Assay. In addition, we demonstrated that pharmacological activation of the PP1 pathway has a negative effect on CSFV replication while inhibition of the PP1 pathway or knockdown of PPP1CB by siRNA had no observed effect. Overall, our data suggests that the CSFV E2 and PPP1CB protein interact in infected cells, and that activation of the PP1 pathway decreases virus replication.
经典猪瘟病毒(CSFV)E2 蛋白是主要的病毒结构糖蛋白,是病毒包膜的重要组成部分。E2 参与病毒的吸附,诱导保护性免疫反应,对猪的毒力至关重要。我们利用酵母双杂交系统鉴定了蛋白磷酸酶 1 催化亚基β(PPP1CB),它是蛋白磷酸酶 1(PP1)复合物的一部分,是 E2 的特定结合宿主伴侣。我们进一步通过两种独立的方法证实了 CSFV 感染的猪细胞中这种相互作用的发生:免疫共沉淀和邻近连接分析。此外,我们证明了 PP1 途径的药理学激活对 CSFV 复制有负面影响,而 PP1 途径的抑制或 siRNA 敲低 PPP1CB 则没有观察到这种影响。总的来说,我们的数据表明,CSFV E2 和 PPP1CB 蛋白在感染细胞中相互作用,而激活 PP1 途径会降低病毒复制。