Wang Qian, Li Yanwei, Dong Hong, Wang Li, Peng Jinmei, An Tongqing, Yang Xufu, Tian Zhijun, Cai Xuehui
Division of Swine Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, No.678, Haping street, Xiangfang District, Harbin, 150069, China.
National Engineering Research Center of Veterinary Biologics, Harbin, 150001, China.
Virol J. 2017 Feb 22;14(1):39. doi: 10.1186/s12985-017-0700-1.
The highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) continues to pose one of the greatest threats to the swine industry. M protein is the most conserved and important structural protein of PRRSV. However, information about the host cellular proteins that interact with M protein remains limited.
Host cellular proteins that interact with the M protein of HP-PRRSV were immunoprecipitated from MARC-145 cells infected with PRRSV HuN4-F112 using the M monoclonal antibody (mAb). The differentially expressed proteins were identified by LC-MS/MS. The screened proteins were used for bioinformatics analysis including Gene Ontology, the interaction network, and the enriched KEGG pathways. Some interested cellular proteins were validated to interact with M protein by CO-IP.
The PRRSV HuN4-F112 infection group had 10 bands compared with the control group. The bands included 219 non-redundant cellular proteins that interact with M protein, which were identified by LC-MS/MS with high confidence. The gene ontology and Kyoto encyclopedia of genes and genomes (KEGG) pathway bioinformatic analyses indicated that the identified proteins could be assigned to several different subcellular locations and functional classes. Functional analysis of the interactome profile highlighted cellular pathways associated with protein translation, infectious disease, and signal transduction. Two interested cellular proteins-nuclear factor of activated T cells 45 kDa (NF45) and proliferating cell nuclear antigen (PCNA)-that could interact with M protein were validated by Co-IP and confocal analyses.
The interactome data between PRRSV M protein and cellular proteins were identified and contribute to the understanding of the roles of M protein in the replication and pathogenesis of PRRSV. The interactome of M protein will aid studies of virus/host interactions and provide means to decrease the threat of PRRSV to the swine industry in the future.
高致病性猪繁殖与呼吸综合征病毒（HP-PRRSV）仍然是养猪业面临的最大威胁之一。M蛋白是PRRSV最保守且最重要的结构蛋白。然而，关于与M蛋白相互作用的宿主细胞蛋白的信息仍然有限。
使用M单克隆抗体（mAb）从感染PRRSV HuN4-F112的MARC-145细胞中免疫沉淀与HP-PRRSV的M蛋白相互作用的宿主细胞蛋白。通过液相色谱-串联质谱（LC-MS/MS）鉴定差异表达的蛋白。筛选出的蛋白用于生物信息学分析，包括基因本体论、相互作用网络和富集的KEGG通路。通过免疫共沉淀（CO-IP）验证了一些感兴趣的细胞蛋白与M蛋白相互作用。
与对照组相比，PRRSV HuN4-F112感染组有10条条带。这些条带包含219种与M蛋白相互作用的非冗余细胞蛋白，通过LC-MS/MS被高度可靠地鉴定出来。基因本体论和京都基因与基因组百科全书（KEGG）通路生物信息学分析表明，鉴定出的蛋白可被分配到几个不同的亚细胞定位和功能类别。相互作用组图谱的功能分析突出了与蛋白质翻译、传染病和信号转导相关的细胞通路。通过免疫共沉淀和共聚焦分析验证了两种感兴趣的细胞蛋白——活化T细胞核因子45 kDa（NF45）和增殖细胞核抗原（PCNA）——可与M蛋白相互作用。
鉴定了PRRSV M蛋白与细胞蛋白之间的相互作用组数据，有助于理解M蛋白在PRRSV复制和发病机制中的作用。M蛋白的相互作用组将有助于病毒/宿主相互作用的研究，并为未来降低PRRSV对养猪业的威胁提供方法。
