Supplementary Figure 1: Cryo-EM analysis of assembly of the RAD51 presynaptic and postsynaptic complexes.
From: Cryo-EM structures of human RAD51 recombinase filaments during catalysis of DNA-strand exchange
(a) Purified human RAD51 and mouse Hop2-Mnd1 were analyzed by SDS-PAGE with Coomassie blue staining and by DNA strand exchange assays. The reaction scheme setup is illustrated. “+” stands for the concentration of Hop2-Mnd1 and +, ++, +++ means 100 nM, 200 nM, and 400 nM, respectively. Rad51 concentration was kept constant at 2 μM in all the assays. The mean values ± s.d. from three independent experiments were presented as bar diagram on the right panel. (b) A representative cryo-EM micrograph of presynaptic filaments and a representative 2D class average are shown in the left panel. 3D reconstruction of the presynaptic filament via IHRSR is shown on the right. (c) A representative cryo-EM micrograph of postsynaptic filaments and a representative 2D class average are shown in the left panel. 3D reconstruction of the postsynaptic complex is shown on the right. (d) 3D reconstruction of the RAD51-ssDNA presynaptic complex incubated with Hop2-Mnd1 is shown on the left panel. The difference map between 3D reconstructions of RAD51-ssDNA presynaptic complex with and without Hop2-Mnd1 incubation is shown in the middle and the right panels with inverted contrast separately. The corresponding atomic model of the postsynaptic complex is shown as reference.
