Supplementary Figure 6: Nucleosome binding activities of various constructs used in this study.

(a) EMSA of the nucleosome binding activities of MtSnf2 (463-1116) with the WT interface and five DNA-binding mutants. 2 nM Cy5-labelled 147 bp NCP was mixed with increasing amounts of proteins (in nM). The samples were resolved by electrophoresis on 4.5% native acrylamide gels, and the fluorescent bands were detected by Typhoon Trio+ imager. With higher concentrations of the proteins, the bound NCPs migrate more slowly, suggesting a heterogeneous population of the protein-bound NCP in the solution, and one NCP might bind more than one molecule of the protein. The star “*” sign indicates the position of free DNA. The bound fractions were quantified as the disappearance of free NCP relative to the intensity of the whole lane, and shown in Fig. 4c. (b) EMSA of nucleosome binding of three constructs of ScSnf2. The quantification of the bound fractions was showed in Fig. 5a. (c) EMSA of nucleosome binding of MtSnf2 (438-1116). EMSA of MtSnf2 (461-1116) was shown in Supplementary Fig. 6a, the left panel (WT interface). Quantification of the bound fractions is shown (right panel). The apparent Kds of the catalytic core MtSnf2 (463-1116) and the HSA-containing MtSnf2 (438-1116) are about 150 nM and 190 nM, respectively. (d) EMSA of nucleosome binding of MtSnf2 with truncation of the C-terminal flexible tail at different positions. EMSA of MtSnf2 (461-1116) was shown in Supplementary Fig. 6a, the left panel (WT interface). The quantification of the bound fractions is showed in Fig. 6c.