Fig. 5

RAD52 prevents fork reversal by SMARCAL1. a, b Cartoon depiction of the DNA substrates and the reversal reaction employing synthetic fork with a 30 nt gap on the leading (a) and lagging (b) strand, respectively. The lengths of the dsDNA and ssDNA features are marked in grey. Green circles depict Cy3 dyes, red circles depict Cy5 dyes. The substrate and the products of the fork reversal reactions are separated on the agarose gels after deproteinization. All reactions were carried out in the presence of 3 nM of forked DNA. c–e Representative gels and quantification of the fork reversal reactions by 20 nM SMARCAL1 in the presence of increasing concentrations of RAD52. Reactions were initiated by addition of SMARCA1 and stopped at 15 min. Only Cy5 channel data were used for quantification. f Representative fork reversal time courses for the DNA substrate containing a leading strand gap. Solid faint lines behind the experimental data indicate fits to a single exponential. g The rates of the fork reversal reactions and their extents were calculated by fitting the data to exponential decay functions as described in the Methods section. The resulting rates are shown for three independent experiments. h, i The same as f, g, but for the substrate with a lagging strand gap. Source data are provided as a Source Data file