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: The primary function of this protein is to encapsidate the viral RNA genome, and it is also thought to participate in the modulation of host cell biology and recruitment of cellular factors to facilitate virus infection.In order to the better understand these latter roles the cellular interactome of PRRSV N protein was defined using label free quantitative proteomics.This identified several cellular factors that could interact with the N protein including poly [ADP-ribose] polymerase 1 (PARP-1), a cellular protein, which can add adenosine diphosphate ribose to a protein.
Affiliation: College of Life Sciences, Northwest A&F University, Yangling, China.Show MeSH
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Mentions: There are several stages to a virus life cycle, and the plaque assay provided a measure of the amount of virus being released into the supernatant. To investigate whether the PARP-1 inhibitor 3-AB affected virus biology in the cell, the abundance of viral proteins were compared using western blot at 24, 48 and 72 h post-infection between treated and untreated PRRSV-infected cells (Fig. 6A). The data indicated that at 24, 48 and 72 h post-infection nsp2, GP2, GP4, GP5, M and N proteins appeared less abundant in 3-AB treated cells than the non-treated cells (Fig. 6A). Western blot analysis indicated that in untreated and uninfected cells, PARP-1 was present in its native state. However, in cells treated with 20 mM 3-AB, PARP-1 is present as a cleavage product as well as native protein (Fig. 6B). This is consistent with the cell viability assay and that PARP-1 is cleaved during apoptosis. PARP-1 is present in the native and cleaved state in infected and treated cells at 24, 48 and 72 h post-infection. However, in untreated cells, PARP-1 is present in the native state at 24 h post-infection and in the native and cleaved state at 48 and 72 h post-infection. This is consistent with cleavage of PARP-1 previously reported in PRRSV infected cells (Lee and Kleiboeker, 2007).
Affiliation: College of Life Sciences, Northwest A&F University, Yangling, China.