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Figure 7
The conserved molecular mechanism of ORF49KSHV interacting with PARP-1. (a) Interaction with PARP-1. FLAG-tagged ORF49KSHV 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 ORF49KSHV with PARP-1. The ORF49KSHV 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 ORF49KSHV. GFP-tagged ORF49KSHV 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 ORF49KSHV mutant. FLAG-tagged ORF49KSHV or ORF49KSHV 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.

IUCrJ
Volume 5| Part 6| November 2018| Pages 866-879
ISSN: 2052-2525
https://doi.org/10.1107/S2052252518013854