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ORIGINAL RESEARCH article
Front. Microbiol.
Sec. Virology
Volume 16 - 2025 | doi: 10.3389/fmicb.2025.1559840
Effect of the CSFV NS5A protein on key proteins in the MAPK and PI3K-mTOR signalling pathways in porcine macrophages
Provisionally accepted
Yidan Wang Xiaoru Zheng Yingying Yang Min Li 
Xinru Zhao 
Juan Huang Qiaoya Zhang Xiaobing Qin 
Ying Yu *
Qing Pan *
Zhi Cao *- College of Veterinary Medicine, Qingdao Agricultural University, Qingdao, China
Classical swine fever (CSF) is a highly contagious disease caused by classical swine fever virus (CSFV). NS5A, a nonstructural protein of CSFV, plays an important role in regulating viral replication and protein translation. The purpose of this study was to investigate the effects of the CSFV NS5A protein on key proteins in the mitogen-activated protein kinase (MAPK) and phosphoinositide 3-kinase (PI3K)-mechanistic target of rapamycin (mTOR) pathways in porcine macrophages. In this study, an NS5A lentivirus was constructed, and 3D4/21 cells were infected. The cells infected for 48 hours were collected for proteomic analysis to screen the differential proteins in the two signalling pathways in the NS5A/control group, and the expression levels of key proteins were verified by Western blotting (Wb). CSFV NS5A lentivirus was successfully constructed and used to infect porcine macrophages, and 23 upregulated proteins and 16 downregulated proteins were found in the MAPK signalling pathway, whereas 5 upregulated and 15 downregulated proteins were found in the PI3K‒mTOR signalling pathway. The results revealed that with increasing infection time, the expression of IKBKG, AKT1, CDC37, MAP3K2 and PKN2 decreased, whereas the expression of MAP3K7 and KRAS2 increased. The 3D4/21 cells infected with NS5A lentivirus and classical swine fever virus were inoculated, and the differential protein expression was verified via Wb. With increasing time, the protein expression levels of IKBKG and KRAS2 increased, whereas the protein expression levels of MAP3K7, MAP3K2, AKT1, CDC37 and PKN2 decreased. This study provides data for revealing the mechanism by which CSFV evades host antiviral immune clearance and has important scientific significance and potential application value.
Keywords: CSFV NS5A, Lentivirus, MAPK pathway, PI3K-mTOR pathway, Differential protein
Received: 13 Jan 2025; Accepted: 11 Feb 2025.
Copyright: © 2025 Wang, Zheng, Yang, Li, Zhao, Huang, Zhang, Qin, Yu, Pan and Cao. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Ying Yu, College of Veterinary Medicine, Qingdao Agricultural University, Qingdao, China
Qing Pan, College of Veterinary Medicine, Qingdao Agricultural University, Qingdao, China
Zhi Cao, College of Veterinary Medicine, Qingdao Agricultural University, Qingdao, China
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