Extended Data Fig. 4: Structural features and functional analysis of BCDX2 subunit active sites.
From: Structural insights into BCDX2 complex function in homologous recombination
a, Overview of the BCDX2 structure shown as a semitransparent cartoon with. ATP and BCDX2 residues contacting ATP are shown as solid sticks (center). Active sites are boxed. Magnified views of BCDX2 active sites are shown in the left and right panels. b, Structure-based sequence alignment of ATP binding sites of BCDX2 subunits and RAD51. Nucleotides are buried at the interface between subunits and the primary ATP binding site is defined as the subunit with its Walker A motif contacting ATP while the secondary ATP binding site is defined as the subunit contributing interacting residues on the other side of ATP. Interacting residues from primary and secondary ATP binding sites are colored green and pink, respectively. RAD51 (PDB: 7EJC) is shown for reference. c, ATPase assays of 1μM indicated BCDX2 variants in the presence or absence of ssDNA after 60 min incubation. Pi indicates released inorganic phosphate after hydrolysis. The histograms show results from three independent experiments, data are presented as mean values ±SD and p-values for the significance of differences in mean values were calculated by the two-sided t-test are indicated. d. ATP hydrolysis with 1 μM WT and indicated mutant BCDX2 complexes with or without ssDNA were assessed for indicated time points and measurements for three independent experiments were plotted as mean values ±SD. ATPase images in panels c and d were derived from same experiment and processed in parallel.
