Detection of interactions between nucleosome arrays mediated by specific core histone tail domains

Abstract

The core histone tail domains play important roles in different stages of chromatin condensation. The tails are required for folding nucleosome arrays into secondary chromatin structures such as the ∼30 nm diameter chromatin fiber and for mediating fiber–fiber interactions important for formation of tertiary chromatin structures. Crosslinking studies have demonstrated that inter-nucleosomal tail–DNA contacts appear in conjunction with salt-induced folding of nucleosome arrays into in higher order chromatin structures. However, since both folding of nucleosome arrays and fiber-fiber interactions take place simultaneously in >2–3 mM MgCl 2 such inter-nucleosome interactions may reflect short range (intra-array) or longer range (inter-array) interactions. Here, we describe a novel technique to specifically identify inter-array interactions mediated by the histone tail domains. In addition, we describe a new method for the preparation of H3/H4 tetramers.