FANCJ-A349P is compromised in its ability to displace DNA-bound protein. (A) The indicated concentrations of FANCJ-WT (upper) or FANCJ-A349P (lower) were incubated with 2mM ATP and DNA substrate (0.5nM) that had streptavidin bound to the covalently linked biotin moiety residing 52 nt from the 5′ end of the radiolabeled oligonucleotide (supplemental Table 1). (B) Quantitative analyses of FANCJ streptavidin displacement assays as shown in panel A. Data represent the mean of at least 3 independent experiments with SD indicated by error bars. (C) Experimental scheme to examine FANCJ displacement of RAD51 protein filament on ssDNA. (D) The degradation of the intact ³²P-labeled ssDNA fragment by exonuclease VII was analyzed in a 10% polyacrylamide gel. The preformed RAD51-ssDNA complex was incubated with FANCJ-WT (lanes 4-7) or FANCJ-A349P (lanes 9-12) followed by the addition of exonuclease VII. Protein concentrations are indicated at the top of the gel. The ssDNA fragment before and after the treatment with exonuclease VII is shown in lanes 1 and 2, respectively. (E) The data from panel D represented as a graph.