Figure 2
From: Human Rad51 mediated DNA unwinding is facilitated by conditions that favour Rad51-dsDNA aggregation
RAD52 stimulates RAD51 mediated dsDNA unwinding. Relaxed ĻX174 DNA (30 Ī¼M) along with Topo I was incubated with RAD51 (3.0 Ī¼M) in presence of increasing concentration of RAD52 (0 Ī¼M, 0.5 Ī¼M, 1.0 Ī¼M & 2.0 Ī¼M) (lanes 1ā4 in each Panel) in binding buffer R at 37Ā°C for 12 minutes, in presence of either ATP, ADP, AMP-PNP, ATPĪ³S, or AMP (1 mM each) (Panels A-E) or no nucleotide (Panel F), followed by deproteinization and agarose gel assay (Methods). Lane 5 in all the Panels has all the components except RAD51. Panel G represents only RAD52 control in presence of ATP. Panel H represents no protein control. Percentage unwound DNA (normalized and quantified) was expressed as a ratio of the intensity associated with unwound DNA band to the sum of intensities corresponding to unwound as well as relaxed dsDNA bands. This was plotted as a function of RAD52 concentration (Panel I). Data points were statistically analyzed as explained earlier.