FIGURE 11.

RAD54-K189R forms more stable complexes with dsDNA than RAD54. a, schematic of the dsDNA dissociation assay. The asterisk indicates the ³²P-label at the DNA 5′ end. b, 16.5 nm RAD54 or RAD54 K189R was incubated with 2% DMSO or 50 μm SN for 5 min at 30 °C. ³²P-labeled pUC19 dsDNA fragments (1313 bp and 1373 bp, 2 μm, nts) were added to form protein-DNA complexes for 10 min. Addition of identical but unlabeled dsDNA fragments (148 μm, nts) initiated the exchange of RAD54 and RAD54 K189R between dsDNA molecules after the protein-DNA complexes were cross-linked by 0.25% glutaraldehyde and analyzed by electrophoresis in 1.5% agarose gels. c, the data presented as a graph. Error bars represent the S.E.