HISTONE CONTRIBUTIONS TO THE STRUCTURE OF DNA IN THE NUCLEOSOME

We describe the application of the hydroxyl radical footprinting technique to examine the contribution of the core histone tails and of histones H-3 and H-4 to the structure of DNA in the nucleosome. We first establish that, as was previously determined for a nucleosome containing a unique sequence of DNA, mixed-sequence nucleosomes contain two distinct regions of DNA structure. The central three turns of DNA in the nucleosome have a helical periodicity of almost-equal-to 10.7 base pairs per turn, while nanking regions have a periodicity of almost-equal-to 10.0 base pairs per turn. Removal of the histone tails does not change the hydroxyl radical cleavage pattern in either mixed- or unique-sequence nucleosome samples. A tetramer of histones H3 and H4, (H3/H4)2, organizes the central 120 base pairs of DNA identically to that found in the nucleosome. Moreover, "tailless" octamers and the (H3/H4)2 tetramer recognize the same nucleosome positioning signals as the intact octamer.