Extended Data Fig. 1: Supporting data for cryo-EM investigation of S. cerevisiae dsDNA-bound replisome:SCF^Dia2 complexes.

a, Schematic of reconstitution approach used for preparation of cryo-EM sample representing terminating SCF^Dia2-bound replisome complexes after translocation onto dsDNA. A schematic of the DNA substrate used is shown in orange, with the 20 nt tract of methylphosphonate (MEP) linkages coloured red. b, Silver-stained SDS-PAGE gels analysing 100 μL fractions taken across 10-30% glycerol gradients, either lacking (top) or containing (bottom) crosslinking agents. Fractions 13+14 used for cryo-EM sample preparation are indicated. * = Cdc34-Ub; ** = Cdc34. Similar results were observed for three independent sample preparations. c, Representative cryo-EM micrograph. d, Representative 2D class averages, 40 nm box width. e, Representative angular distribution of particle orientations. A correspondingly oriented model is shown to the right for reference. f, Fourier shell correlation graphs for maps used in model building. The resolution of reconstructions calculated at the FSC=0.143 criterion are reported in Extended Data Fig. 2. g, Model-to-map correlation graphs. h, Cryo-EM density maps relevant to model building, coloured by local resolution. For gel source data, see Supplementary Fig. 1.