Borca Manuel V, O'Donnell Vivian, Holinka Lauren G, Rai Devendra K, Sanford Brenton, Alfano Marialexia, Carlson Jolene, Azzinaro Paul A, Alonso Covadonga, Gladue Douglas P
Agricultural Research Service and Department of Homeland Security, Plum Island Animal Disease Center, Greenport, NY 11944, USA.
Agricultural Research Service and Department of Homeland Security, Plum Island Animal Disease Center, Greenport, NY 11944, USA; Departments of Pathobiology and Veterinary Science, University of Connecticut, Storrs, CT 06269, USA.
Virus Res. 2016 Sep 2;223:181-9. doi: 10.1016/j.virusres.2016.07.013. Epub 2016 Aug 3.
African swine fever virus (ASFV) is the etiological agent of a contagious and often lethal disease of domestic pigs that has significant economic consequences for the swine industry. The viral genome encodes for more than 150 genes, and only a select few of these genes have been studied in some detail. Here we report the characterization of open reading frame Ep152R that has a predicted complement control module/SCR domain. This domain is found in Vaccinia virus proteins that are involved in blocking the immune response during viral infection. A recombinant ASFV harboring a HA tagged version of the Ep152R protein was developed (ASFV-G-Ep152R-HA) and used to demonstrate that Ep152R is an early virus protein. Attempts to construct recombinant viruses having a deleted Ep152R gene were consistently unsuccessful indicating that Ep152R is an essential gene. Interestingly, analysis of host-protein interactions for Ep152R using a yeast two-hybrid screen, identified BAG6, a protein previously identified as being required for ASFV replication. Furthermore, fluorescent microscopy analysis confirms that Ep152R-BAG6 interaction actually occurs in cells infected with ASFV.
非洲猪瘟病毒(ASFV)是家猪一种具有传染性且往往致命疾病的病原体,对养猪业造成重大经济影响。该病毒基因组编码超过150个基因,其中只有少数几个基因得到了较为详细的研究。在此,我们报告了具有预测补体控制模块/SCR结构域的开放阅读框Ep152R的特征。该结构域存在于痘苗病毒蛋白中,这些蛋白在病毒感染期间参与阻断免疫反应。构建了携带Ep152R蛋白HA标签版本的重组ASFV(ASFV-G-Ep152R-HA),并用于证明Ep152R是一种早期病毒蛋白。构建缺失Ep152R基因的重组病毒的尝试一直未成功,这表明Ep152R是一个必需基因。有趣的是,使用酵母双杂交筛选对Ep152R的宿主蛋白相互作用进行分析,鉴定出BAG6,一种先前被确定为ASFV复制所必需的蛋白。此外,荧光显微镜分析证实Ep152R与BAG6的相互作用确实发生在感染ASFV的细胞中。