Extended Data Fig. 5: BRCA2 protects RAD51 filaments from dismantling by FBH1 and RADX.
From: BRCA2 prevents PARPi-mediated PARP1 retention to protect RAD51 filaments
Normalized FRET histograms of (a) DNA only (first panel), RAD51 only (second panel), RAD51 with FBH1 (third panel) (n = 320), BRCA2 with RAD51(fourth panel) and finally BRCA2-RAD51 with FBH1 (fifth panel) (n = 276) (b) DNA only (first panel), RAD51 only (second panel), RAD51 with RADX (third panel) (n = 446), BRCA2 with RAD51(fourth panel) and finally BRCA2-RAD51 with RADX (fifth panel) (n = 391) (c) DNA only (first panel), RAD51 only (second panel), RAD51 with PARP1 + PARPi (third panel), RAD51 with PARP1 + PARPi + FBH1 (n = 240) (fourth panel),RAD51 with PARP1 + PARPi + RADX (n = 220) (fifth panel) and BRCA2-RAD51 with PARP1 + PARPi (sixth panel). (d) Effect of different replication fork proteins that governs fork stability such as RADX and FBH1 on RAD51 filament stability, cartoon depiction on the right, defining the specificity that PARPi have on PARP1 mediated changes to RAD51 filaments. Data shown is mean ± s.d (r = 2). The number of analyzed molecules and replicates are denoted by n and r, respectively. Data for DNA only, DNA with RAD51 and DNA with BRCA2-RAD51 was replotted here for comparison purposes. Cartoons were created in BioRender. Lahiri, S. (2025) https://biorender.com/l07t264. [BRCA2] = 0.04 µM | [RAD51] = 0.4 µM | [ATP] = 2 mM. [RADX] = 0.04 µM | [FBH1] = 0.04 µM. [PARP1] = 0.4 µM | NAD+ = 1 mM | [PARPi] = 0.1 µM talazoparib (BMN673).
