Extended Data Fig. 3: Kinetic analysis of BRCA2-RAD51 binding on resected DNA in presence of PARP1 + PARPi.
From: BRCA2 prevents PARPi-mediated PARP1 retention to protect RAD51 filaments
(a-b) A TDP generated from HMM analysis of the smFRET trajectories for RAD51 and PARP1 + PARPi (a) bound to 3′ tailed DNA highlighting the transition densities corresponding to each state (PR + N, IE, E). From the TDP, individual states were selected, and the corresponding dwell-time histograms (bin size = 90 msec) were fitted using a single exponential function to obtain the apparent rates of transitions out of the states along with the FRET population histograms for unprocessed and unnormalized raw data from ebFRET output (b) (n = 850; r = 3). (c-d) A TDP generated from HMM analysis of the smFRET trajectories for BRCA2-RAD51 and PARP1 + PARPi (c) bound to 3′ tailed DNA with transition states (N′, IE′, E′) and corresponding dwell-time histograms (bin size = 90 msec) along with the FRET population histograms for unprocessed and unnormalized raw data from ebFRET output (d) (n = 652; r = 4). Brighter colors represent more frequent transitions, and the frequency scale is shown to the right of the graphs. 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. [PARP1] = 0.4 µM | NAD+ = 1 mM | [PARPi] = 0.1 µM talazoparib (BMN673).
