Journal
VIRUSES-BASEL
Volume 14, Issue 5, Pages -Publisher
MDPI
DOI: 10.3390/v14050863
Keywords
Senecavirus A (SVA); PK-15 cells; tandem mass tags (TMT); liquid chromatography-tandem mass spectrometry (LC-MS; MS); differentially expressed proteins (DEPs); Mx1
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Funding
- Beijing Natural Science Foundation [6212013]
- National Natural Science Foundation of China [32072840]
- China Agriculture Research System of MOF and MARA [CARS-35]
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This study analyzed the dynamic changes in the proteome of PK-15 cells during Senecavirus A (SVA) infection using tandem mass tags (TMT) labeling coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS). The results showed significant differential expression of proteins at different time points of infection, with these proteins mainly involved in host immune responses and metabolic processes. In addition, the study demonstrated that Mx1 protein exerts antiviral activity against SVA by interacting with VP1 and VP2 proteins.
As an emergent picornavirus pathogenic to pigs, Senecavirus A (SVA) can replicate in pig kidneys and proliferates well in porcine kidney epithelial PK-15 cells. Here, tandem mass tags (TMT) labeling coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to analyze the proteome dynamic changes in PK-15 cells during SVA infection. In total, 314, 697 and 426 upregulated differentially expressed proteins (DEPs) and 131, 263 and 342 downregulated DEPs were identified at 12, 24 and 36 hpi, respectively. After ensuring reliability of the proteomic data by quantitative PCR and Western blot testing of five randomly selected DEPs, Mx1, eIF4E, G6PD, TOP1 and PGAM1, all the DEPs were subjected to multiple bioinformatics analyses, including GO, COG, KEGG and STRING. The results reveal that the DEPs were mainly involved in host innate and adaptive immune responses in the early and middle stages of SVA infection, while the DEPs mainly participated in various metabolic processes in the late stage of infection. Finally, we demonstrated that Mx1 protein exerts antiviral activity against SVA by interacting with VP1 and VP2 proteins dependent on its GTPase, oligomerization and interaction activities, while Mx1 interacts with VP3 only depending on its oligomerization activity. Collectively, our study provides valuable clues for further investigation of SVA pathogenesis.
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