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. Author manuscript; available in PMC: 2018 Aug 13.
Published in final edited form as: ACS Chem Biol. 2017 Oct 19;12(11):2866–2874. doi: 10.1021/acschembio.7b00445

Figure 4. GeA-69 engages PARP14 MD2 and prevents localisation to the sites of DNA damage.

Figure 4

(A) PARP14 MD2 wild-type is recruited to sites of laser-induced DNA damage in contrast to the corresponding ADPR binding deficient mutant G1044E. U-2 OS cells were transfected with YFP-PARP14 MD2 wt or G1044E mutant, subjected to laser micro-irradiation (white arrows) and imaged at the indicated times. (B) PARP14 MD2 recruitment depends on the presence of PARP1. U-2 OS/PARP1+/+ and U-2 OS/PARP1-/- cells were transfected with YFP-PARP14 MD2 wt, subjected to laser micro-irradiation and imaged at the indicated times. (C) PARP14 MD2 recruitment depends on the activity of PARP1. U-2 OS cells transiently expressing YFP-PARP14 MD2 wt were pre-treated for 1 hour with DMSO or with 10 µM of the PARP1 inhibitor Olaparib, subjected to laser micro-irradiation and imaged at the indicated times. (D) GeA-69 prevents recruitment of PARP14 MD2 to sites of laser-induced DNA damage. U-2 OS cells transiently expressing YFP-PARP14 MD2 or full-length YFP-ALC1 were pre-treated with DMSO, GeA-69 or the negative control inhibitor MnK2-68, subjected to laser micro-irradiation and imaged at the indicated times.