Extended Data Fig. 1: Kinetic analysis of BRCA2 modulating distinct states of RAD51 filament.
From: BRCA2 prevents PARPi-mediated PARP1 retention to protect RAD51 filaments
(a) Autoradiogram of DNA strand exchange reaction with 3′ tailed DNA and increasing concentrations of RAD51 in the presence of ATP. (b) A pseudo-3D plot, Transition Density plot (TDP), generated from HMM analysis of the smFRET trajectories for RAD51 only bound to 3′ tailed DNA highlighting the transition densities corresponding to each state (N, IE, E). (c) Dwell time histograms (bin size = 90 msec) for each of the FRET states as obtained from HMM analysis (Supplementary Table 1) and FRET population histograms for unprocessed and unnormalized raw data from ebFRET output (n = 891; r = 3). (d-e) TDP generated from HMM analysis of the smFRET trajectories for BRCA2-RAD51 bound to 3′ tailed DNA highlighting the transition densities corresponding to each state (N′, IE′, E′) along with dwell time histograms (bin size = 90 msec) for each of the FRET states as obtained from HMM analysis (Supplementary Table 1) (n = 670; r = 3). In both cases, individual states were selected from the TDP, and the corresponding dwell-time histograms were fitted using a single exponential function to obtain the apparent rates of transitions out of the states The number of analyzed molecules and replicates are denoted by n and r, respectively. [BRCA2] = 0.04 µM | [RAD51] = 0.4 µM | [ATP] = 2 mM.
