Fig. 1

SHPRH is a dsDNA and a nucleosome-binding protein. a Schematic representation of the SHPRH protein domains (see main text for details). b SHPRH, SNF2H and the human SWI/SNF complex (~ 12 subunits) preparations which were used in assays were separated by SDS PAGE and stained with Coomassie blue. c Nucleosome electrophoretic mobility shift assay. 20 nM of radioactively labeled 147-bp DNA (lanes 1–4), the same DNA assembled into nucleosome core particles (lanes 5–8), a 227-bp DNA (lanes 9–12) or the same DNA assembled into mononucleosomes (Nuc + 80-bp of extranucleosomal DNA; lanes 13–16) were incubated with increasing concentrations of SHPRH (57, 113, 170 nM). Free or SHPRH-bound DNAs and nucleosomes were separated by native PAGE and visualized using a phosphorimager screen. The star indicates minor non-specific PCR-products. Positions of free DNA and nucleosomes (Nuc) are indicated on the right. d Quantification of the nucleosome electrophoretic mobility shift assay shown in (c). Percentages of the unbound DNA or nucleosome probes relative to the respective (−) SHPRH lanes (Y-axis) were plotted as a function of SHPRH concentration, as indicated on the X-axis. Error bars reflect standard deviation derived from two independent experiments. Note that error bars may not be visible for data with very small standard deviations