FIG. 5.

Both Isw2p and Itc1p are required for biochemical activities of Isw2 complex. (A) ATPase assays using native Isw2 complex, monomeric Isw2p, monomeric Itc1p, and reconstituted Isw2 complex. Assays were done in the presence of buffer (B) or immobilized nucleosomal arrays (N). (B) Chromatin remodeling assay performed on preassembled nucleosomal arrays on immobilized templates. After incubation of the templates with ATP and the fractions indicated above, limited (lanes 1, 3, 5, 7, and 9) and extensive (lanes 2, 4, 6, 8, and 10) MNase digestion was performed. Nucleosomal arrays containing 25 ng of DNA and 45 fmol of Isw2 complex, Isw2p, or Itc1p were used. Arrowheads denote mononucleosome signals increased by native and reconstituted Isw2 complex. The arrow on the right indicates a dinucleosome signal that is shifted upward by the native Isw2 complex (lane 3). DNA was visualized by Southern blotting.