Figure 8. Mutation of AcidicN, the AcidicN binding interface or AutoN suppresses dependence on the H4-tail.
(A) H4-tail dependence of the remodeling activities of ISWI variants. Values were calculated from the observed remodeling rate constants obtained for WT and tail-less H4 chromatin (Figure 8—figure supplement 1E). (B) ATP hydrolysis measurements of ISWT+6, ISWI2RA and ISWI+6; 2RA in absence or presence of saturating concentrations of DNA and chromatin.
Figure 8—figure supplement 1. Raw data of the remodeling assays.
(A–D) Determination of rate constants (kobs) from remodeling assays for ISWI+6, ISWI2RA and ISWI+6; 2RA. Shown are exemplary time courses on nucleosomal arrays containing wild-type (A,C) and tail-less H4 (B,D). Data were fit to a single exponential function to extract the rate constant kobs. (E) Rate constants for remodeling of ISWI variants used in this study (all 300 nM). Errors are s.d. (n ≥ 3) for ISWIWT, ISWIΔppHSA, and ISWI+6 and minimal and maximal values of two independent measurements for all other variants. Samples, in which the enzyme concentration was not saturating, are indicated (§). Data marked with asterisks (*) were replotted from previous figures for better overview.