Porcine epidemic diarrhea virus (PEDV) is a coronavirus that causes severe diarrhea in pigs of all ages and a high fatality rate in neonates. The PEDV membrane protein (M) plays crucial roles in viral assembly, viral budding and host immune regulation, most likely by interacting with host cell proteins that have yet to be identified. In this study, co-immunoprecipitation (Co-IP) using an M-specific monoclonal antibody, coupled with LC-MS/MS, was employed to identify M protein-interacting proteins in PEDV-infected cells. Three viral proteins (S, E and ORF3) and 218 host cell proteins were identified as putative M-interacting partners. Bioinformatic analysis showed that the identified host cell proteins were related to 131 signal pathways and 10 biological processes. In addition, interaction between translation initiation factor 3(eIF3L) and M protein was validated by Co-IP. Down-regulation of eIF3L expression significantly increased viral production, which suggests that eIF3L could be a negative regulator in PEDV replication. This interactome study of the PEDV M protein will serve to clarify its function during viral replication.

猪流行性腹泻病毒（PEDV）是一种冠状病毒，可引起所有年龄段的猪严重腹泻，新生儿死亡率高。PEDV膜蛋白（M）在病毒装配，病毒出芽和宿主免疫调节中起关键作用，最有可能通过与尚未鉴定的宿主细胞蛋白相互作用而发挥作用。在这项研究中，使用M特异性单克隆抗体与LC-MS / MS的共免疫沉淀（Co-IP）来鉴定PEDV感染细胞中与M蛋白质相互作用的蛋白质。三种病毒蛋白（S，E和ORF3）和218个宿主细胞蛋白被确定为可能的M相互作用伴侣。生物信息学分析表明，鉴定出的宿主细胞蛋白与131个信号通路和10个生物学过程有关。此外，Co-IP验证了翻译起始因子3（eIF3L）与M蛋白之间的相互作用。eIF3L表达的下调显着增加了病毒的产生，这表明eIF3L可能是PEDV复制中的负调节剂。PEDV M蛋白的这一相互作用组研究将有助于阐明其在病毒复制过程中的功能。