Figure 5. RPA plays a key role in resection initiation by both the Dna2- and CtIP-dependent pathways. (A) Effects of RPA depletion on the generation of short resection initiation products in the extract. (B) Effects of RPA depletion on the generation of long resection initiation products in the Dna2-depleted extract. (C) Purified recombinant Flag-Dna2(WT) and Flag-Dna2(27–1053) expressed in insect cells. (D) Comparison of the flap endonuclease activity of Flag-Dna2(WT) and Flag-Dna2(27–1053) in vitro. (E) Co-immunoprecipitation of RPA with Flag-Dna2(WT) and Flag-Dna2(27–1053) added to the extract. (F) Comparison of the ability of Flag-Dna2(WT) and Flag-Dna2(27–1053) to rescue short resection initiation products in the Dna2-depleted extract. (G) Effects of RPA depletion on the binding of NBS1, CtIP, Dna2, Exo1 and PCNA to the DNA substrate in the extract. (H) Effects of RPA depletion on the binding of NBS1, CtIP, Exo1 and PCNA to the DNA substrate in the Dna2-depleted extract. (I) Comparison of the ability of RPA(WT) and RPA(1NΔ) to rescue short resection initiation products in the RPA-depleted extract. (J) Comparison of the ability of RPA(WT) and RPA(1NΔ) to rescue long resection initiation products in the extract depleted of both Dna2 and RPA.
Image published in: Paudyal SC et al. (2017)
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