Abstract
Porcine sapelovirus A (PSV) is a single stranded, positive-sense, non-enveloped RNA virus that causes enteritis, pneumonia, polioencephalomyelitis, and reproductive disorders in pigs. Research on PSV infection and interaction with host cells is unclear. In this study, we applied tandem mass tag proteomics analysis to investigate the differentially expressed proteins (DEPs) in PSV-infected pig kidney (PK)-15 cells and explored the interactions between PSV and host cells. Here we mapped 181 DEPs, including 59 up-regulated and 122 down-regulated DEPs. Among them, osteopontin (SPP1), induced protein with tetratricopeptide repeats 5 (IFIT5), ISG15 ubiquitin-like modifier (ISG15), vinculin (VCL), and syndecan-1 (SDC1) were verified significantly changed using RT-qPCR. Additionally, overexpression of SDC1 promoted PSV viral protein (VP)1 synthesis and virus titer, and silencing of SDC1 revealed the opposite results. Our findings show that SDC1 is a novel host protein and plays crucial roles in regulating PSV replication.
Highlights
Porcine sapelovirus A (PSV), a member of the genus sapelovirus in the family Picornaviridae, is a single-stranded, positive-sense, non-enveloped RNA virus with a full-length genome of 7.5–8.3 nucleotide that contains a 3 poly (A) tail with a variable length from 65 to 100 nt
A(Fdigdurietio1Bn)a. lAdddaittaionoabl tdaaitnaeodbtafrinoemd fWromesWteersntebrnlobtloatnaanalylyssiiss rreevveealaeldedthatthtahte the virus protein (VPvp)irr1outseeixpnrpcorhteeaisnnsg(ieVoonPf)1PleSvVeex-ilpnrwfeescastiesodnhPilgeKvh-e1l5awtcea8lslshh, ipwgihe(acFthio8gshuepr8ie(hF1piCgi ua)sr.etTh1heCet)i.rmTeehfoeprroeeifn,otrtseo,fotborebftuetretttrhereurunpnddroeetrresostatmanindc d protein change of PanSaVly-isnisf.ected pig kidney (PK)-15 cells, we chose 8 hpi as the time points for further proteomic analysis
Data are expressed as mean ± SD for two independently experiments. 2.2
Summary
Porcine sapelovirus A (PSV), a member of the genus sapelovirus in the family Picornaviridae, is a single-stranded, positive-sense, non-enveloped RNA virus with a full-length genome of 7.5–8.3 nucleotide (nt) that contains a 3 poly (A) tail with a variable length from 65 to 100 nt. Identification of host proteins, which viruses use during their replicative cycle, is essential. Investigation of the changed genes or proteins using the proteomics upon virus infection host cells has become an effective instrument for several viral pathogens, including duck hepatitis A virus type 1 [10], Bombyx mori nucleopolyhedrovirus [11], caprine parainfluenza virus type 3 [12], influenza virus [13], and Dengue virus [14]. T2t0in20g, 2a1s,s4a3y86of PSV virus protein (VP) expression in PK-15 cells at the indicated time points (32o–f1126 h). Identification of Differentially Expressed Proteins (DEPs) Protein extracts from mock-infected and PSV-infected samples were subjected to TMT-coupled LC–MS/MPSroatneainlyesixstraancdtsaftrootmal omf o5c6k90-inpfreoctteeindsawnedrePSidVe-nintiffieecdte(dSuspamplpemlesenwtaerrye TsaubbljeecSt1e)d. RPGR (Fragment) Uncharacterized protein Nucleoporin 153 Interleukin enhancer binding factor 3 Syndecan T-complex protein 1 subunit alpha NOVA alternative splicing regulator 2 SET domain containing 2 Uncharacterized protein PHD finger protein 23 Heterogeneous nuclear ribonucleoprotein U Nucleoporin like 2 Syndecan-4 Activating transcription factor 6 (Fragment) Heterogeneous nuclear ribonucleoprotein U Tumor necrosis factor receptor superfamily member 1A Zinc finger CCCH-type containing 14 Heterogeneous nuclear ribonucleoprotein M Olfactory receptor Cleavage stimulation factor subunit 2 tau variant
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