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.
Bottom Line: Porcine reproductive and respiratory syndrome virus (PRRSV) is a major threat to the swine industry and food security worldwide.The nucleocapsid (N) protein is a major structural protein of PRRSV.Use of the PARP-1 small molecule inhibitor, 3-AB, in PRRSV infected cells demonstrated that PARP-1 was required and acted as an enhancer factor for virus biology.
Affiliation: College of Life Sciences, Northwest A&F University, Yangling, China.Show MeSH
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Mentions: Different doses of 3-AB were assessed for potential anti-viral activity by infecting cells with the same MOI of PRRSV and then treating cells with 5, 10, and 20 mM 3-AB over the infection period. Progeny virus production was compared by endpoint dilution assay to an untreated control at 12, 24, 48, 72 and 96 h post-infection. The data indicated that at the 12, 24 and 48 h time points, using a one-way ANOVA test, there was no significant difference (p > 0.05) in progeny virus production, however at 72 and 96 h post-infection there was a significant difference (p < 0.05) in progeny virus production between the treatments compared to the control non-treated infected cells (Fig. 4B). This was also confirmed using indirect immunofluorescence microscopy to detect viral antigens in virus-infected cells at 24, 48, 72 and 96 h post-infection compared to an untreated control (Fig. 5A). There were considerably less foci of infection in the treated cells compared to untreated control infected cells (Fig. 5B) (data shown for the 20 mM concentration of 3-AB). This confirmed the titration data that virus biology was adversely affected by 3-AB, with virus output being reduced to 1–2 log10 at 96 h post-infection compared to 7 log10 for the infected, but untreated control (Fig. 4B).
Affiliation: College of Life Sciences, Northwest A&F University, Yangling, China.