Resolution of the cellular proteome of the nucleocapsid protein from a highly pathogenic isolate of porcine reproductive and respiratory syndrome virus identifies PARP-1 as a cellular target whose interaction is critical for virus biology

Abstract

HighlightsProteomics was used to identify the cellular interactome of PRRSV N protein.The interactome included translation factors and PARP-1.Inhibition of PARP-1 by the small molecule 3-AB resulted in a decrease in virus infection.Sustained treatment of PRRSV infected cells with 3-AB suggested resistance free antiviral activity.

Poly(ADP-Ribose) Polymerases in Host-Pathogen Interactions, Inflammation, and Immunity

“…The nucleocapsid protein of porcine reproductive and respiratory syndrome virus (PRRSV) binds to PARP1, and the interaction is critical for viral replication, since the use of a PARP inhibitor led to inhibited viral growth (156). In an intriguing parallel, Grunewald et al have shown that the nucleocapsid proteins of both ␣and ␤-coronaviruses are MARylated during infection (157).…”

Determination of the interactome of non-structural protein12 from highly pathogenic porcine reproductive and respiratory syndrome virus with host cellular proteins using high throughput proteomics and identification of HSP70 as a cellular factor for virus replication

“…Although several studies have been focused on the relationship between PRRSV viral proteins such as N [17,34], NSP1 [35,36], NSP2 [37] and whole virion [38,39] and the host cellular proteins, cellular interactome of PRRSV NSP12 has not been reported to the best of our knowledge. Here, high throughput proteomics coupled with immunoprecipitated strategy was used to identify and quantify cellular proteins which potentially interacted with PRRSV NSP12 and participated in viral replication,…”

“…The interactome of PRRSV NSP12 was determined by high-affinity GFP pull-down coupled with label free mass spectrometry-based approach, which we have used previously to study virus/host protein interactions [16][17][18][19]. Our data identified112 cellular proteins probably interacted with NSP12, and these proteins were mainly nucleic acid binding proteins or chaperones and were grouped into several distinct functional clusters.…”

“…Third, 293T cells have already been successfully used in many studies to identify the cellular partners for different viral proteins [16][17][18][19]. Previous studies have reported that Marc-145 cells and PAMs behave quite differently in PRRSV infection [47].…”

“…The reaction procedure was 95°C for 10 minutes, followed by 40 cycles at 95°C for 3 seconds then 60°C for 30 seconds. Relative levels of N gene mRNA was calculated using the 2 -ΔΔt method of relative quantification with GAPDH[17]. Specific primers used were as follows: N-3') for the detection of GAPDH mRNA.…”