Extended Data Fig. 7: Binding of replication fork proteins during NSD.
From: Replication fork uncoupling causes nascent strand degradation and fork reversal
(A) Cartoon depicting two different models of CMG helicase behavior during fork reversal and NSD. In (i) the CMG helicase translocates onto double-stranded DNA, as suggested1,14, which would result in its removal from DNA15,16,17,18. In (ii) the replisome remains on DNA, suggesting that it resides in a ssDNA bubble ahead of the reversed fork. (B) In parallel to Fig. 5A chromatin bound proteins were analyzed in the presence of DNA2-i. RPA was detected by Western Blotting. (C) Quantification of RPA signal from Fig. 5B. and (b). Mean ± S.D., n = 3 independent experiments. Between 0 and 60 minutes, when most fork reversal took place (Fig. 3D), most RPA signal was due to DNA2 activity. Thus, most RPA signal during this time was due to DNA2-dependent NSD, which may mask any dissociation of RPA that occurred due to reannealing of parental DNA strands.
