Abstract
Classical swine fever (CSF) is a highly contagious viral disease causing severe economic losses to the swine industry. As viroporins of viruses modulate the cellular ion balance and then take over the cellular machinery, blocking the activity of viroporin or developing viroporin-defective attenuated vaccines offers new approaches to treat or prevent viral infection. Non-structural protein p7 of CSF virus (CSFV) is a viroporin, which was highly involved in CSFV virulence. Deciphering the interaction between p7 and host proteins will aid our understanding of the mechanism of p7-cellular protein interaction affecting CSFV replication. In the present study, seven host cellular proteins including microtubule-associated protein RP/EB family member 1 (MAPRE1), voltage-dependent anion channel 1 (VDAC1), proteasome maturation protein (POMP), protein inhibitor of activated STAT 1 (PIAS1), gametogenetin binding protein 2 (GGNBP2), COP9 signalosome subunit 2 (COPS2), and contactin 1 (CNTN1) were identified as the potential interactive cellular proteins of CSFV p7 by using yeast two-hybrid (Y2H) screening. Plus, the interaction of CSFV p7 with MAPRE1 and VDAC1 was further evaluated by co-immunoprecipitation and GST-pulldown assay. Besides, the p7-cellular protein interaction network was constructed based on these seven host cellular proteins and the STRING database. Enrichment analysis of GO and KEGG indicated that many host proteins in the p7-cellular protein interaction network were mainly related to the ubiquitin-proteasome system, cGMP-PKG signaling pathway, calcium signaling pathway, and JAK-STAT pathway. Overall, this study identified potential interactive cellular proteins of CSFV p7, constructed the p7-cellular protein interaction network, and predicted the potential pathways involved in the interaction between CSFV p7 and host cells.
Highlights
Classical swine fever (CSF), caused by CSF virus (CSFV), is a severe viral disease causing severe economic losses to the swine industry (Paton and Greiser-Wilke, 2003)
The recombinant bait plasmid pGBKT7-p7 was screened against the peripheral blood mononuclear cell (PBMC) cDNA library, and seven proteins were identified as potential binding partners of the CSFV p7 and their potential functions were listed in Table 2 based on the UniProt database
As yeast cells cotransformed with pGADT7-cellular protein and pGBKT7-p7 could grow on three kinds of synthetically defined media and form blue colonies on QDO/X/A medium (Figure 1), it implied that the host cellular protein interacted with CSFV p7 in yeast cells
Summary
Classical swine fever (CSF), caused by CSF virus (CSFV), is a severe viral disease causing severe economic losses to the swine industry (Paton and Greiser-Wilke, 2003). A series of vaccination strategies have been developed to curb the outbreak events, CSF is still posing an ongoing threat to the swine industry. Studies reported that the emergence of CSFV strains with moderate or attenuated virulence could result in persistent recessive infection and immunosuppression in pigs, making it more intractable to suppress and control CSF (Edwards et al, 2000; Moennig, 2000). The persistent research on the pathogenesis of CSF will be beneficial to the development of new vaccines or therapies for effectively controlling CSF. CSFV, belonging to the Pestivirus genus within the Flaviviridae family, is a small, enveloped virus with a 12.3 kb single positivesense RNA genome. Its genome consists of a single large open reading frame (ORF) flanked by a 5’-untranslated region (5’UTR) and a 3’-UTR. The ORF encodes a polypeptide precursor, which could be cleaved into twelve mature proteins, including four structural proteins (C, Erns, E1, and E2) and eight nonstructural proteins (Npro, p7, NS2, NS3, NS4A, NS4B, NS5A, and NS5B) (Thiel et al, 1991)
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