Fig. 5: Inhibition of pFLP repair leads to replisome disassembly during DNA replication. | Nature Communications

Fig. 5: Inhibition of pFLP repair leads to replisome disassembly during DNA replication.

From: PARP1-dependent DNA-protein crosslink repair

Fig. 5

A pCTRL and pFLP were replicated in the presence or absence of [α-32P]dATP and PARPi (licensing in one volume of HSS for 60 min followed by the addition of two volumes of NPE). Samples were retrieved at indicated time points following NPE addition and analyzed by native agarose gel electrophoresis (top radiograph) or western blotting (bottom immunoblots). Samples were immunoblotted for p.CHK1 and loading control (ORC2). B Top, leftward, and rightward fork models of replisome disassembly when encountering the Flp-nick crosslink. Bottom, pFLP was replicated in the presence of [α-32P]dATP and with or without PARPi. Samples were retrieved at indicated time points, phenol-chloroform extracted, digested with PstI and SapI, and resolved on a denaturing polyacrylamide gel. C Schematic of pFLP linearization by ScaI and the potential DNA species occurring on a 2D gel upon plasmid replication. D Indicated samples from (B) were linearized by ScaI and run in two dimensions. Source data are provided as a Source Data file.

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