Fig. 3

SHPRH does not display conventional nucleosome remodeling activity. a Nucleosome mobilization assay. Increasing concentrations of enzymes (SNF2H: 13 nM, 26 nM, 52 nM or SHPRH: 43 nM, 85 nM, 170 nM) were incubated with 20 nM of radiolabeled end-positioned nucleosome comprising an 80-bp DNA overhang. Migration of the end- or the centrally positioned-nucleosome (filled circle) are indicated on the left. b Analysis of nucleosome remodeling activities by restriction enzyme accessibility (REA) assay. The assay measured the ability of the tested enzymes to expose an MfeI restriction site in the nucleosome at + 28-bp. Control (buffer only), remodeling factors (hSWI/SNF 93 ng and 186 ng in 20 μl; SHPRH 17 nM, 34 nM and 68 nM) were incubated with (20 nM) radiolabeled nucleosomes with a 80-bp DNA overhang, in the presence of ATP (2 mM) and the reaction were stopped at 20, 40 and 60 min