Figure 7.

The conserved molecular mechanism of ORF49_KSHV interacting with PARP-1. (a) Interaction with PARP-1. FLAG-tagged ORF49_KSHV was transfected into HEK293T cells and incubated for 48 h. The cells were harvested and analyzed by co-IP assays with an anti-FLAG antibody. (b) SPR analysis of ORF49_KSHV with PARP-1. The ORF49_KSHV protein was injected at six concentrations (7.12, 3.56, 1.78, 0.89, 0.44 and 0.22 µM). Dissociation data were collected for 120 s. The black lines show the actual data; the orange lines are curve fits. (c, d) Inhibition of interactions between RTA and PARP-1 by ORF49_KSHV. GFP-tagged ORF49_KSHV was co-transfected with FLAG-tagged RTA into HEK293T cells. The cells were harvested 48 h post-transfection and subjected to co-IP assays with an anti-FLAG antibody (c) or an anti-PARP-1 antibody (d). The results were analyzed by Western blotting. (e) PARP-1 interaction of the ORF49_KSHV mutant. FLAG-tagged ORF49_KSHV or ORF49_KSHV ΔN was transfected into HEK293T cells. The cells were harvested 48 h post-transfection and assayed for PARP-1 interaction by co-IP assays with an anti-PARP-1 antibody. The results were analyzed by Western blotting.