Fig. 4.

The effect of DNA and the impact on NCP dynamics. a Superimposed ¹H,¹⁵N HSQC spectra of CHD4 PHD1/2 recorded with and without tetrasome. b Crystal structure of the NCP (PDB ID: 1KX5) with H3 (cyan ribbon), Cy5 at H2AK119C (purple circle), Cy3 at the 5 prime end of 601 L (orange circle), and the LexA target site (brick red) are shown and labeled. c FRET efficiency (E) of the Cy3-Cy5 labeled nucleosomes with increasing concentration of LexA (0–10 μM). LexA binds to its target site and traps the nucleosome in a partially unwrapped state, resulting in a decrease in FRET as [LexA] increases. The LexA titration data were fit to a binding isotherm with an S _1/2 = 1.4 μM. (d–f) FRET efficiencies of the Cy3-Cy5-NCP upon titration with CHD4 PHD1/2, PHD1 or PHD2 in the presence of 1.4 μM of LexA. Error bars represent SD based on three experiments