Temperature and osmotic stress dependence of the thermodynamics for binding linker histone H1 0 , Its carboxyl domain (H1 0 -C) or globular domain (H1 0 -G) to B-DNA

Abstract

Linker histones (H1) are the basic proteins in higher eukaryotes that are responsible for the final condensation of chromatin. In contrast to the nucleosome core histone proteins, the role of H1 in compacting DNA is not clearly understood. In this study ITC was used to measure the binding constant, enthalpy change, and binding site size for the interactions of H10, or its C-terminal (H10-C) and globular (H10-G) domains to highly polymerized calf-thymus DNA at temperatures from 288 K to 308 K. Heat capacity changes, ΔCp, for these same H10 binding interactions were estimated from the temperature dependence of the enthalpy changes. The enthalpy changes for binding H10, H10-C, or H10-G to CT-DNA are all endothermic at 298 K, becoming more exothermic as the temperature is increased. The ΔH for binding H10-G to CT-DNA is exothermic at temperatures above approximately 300 K. Osmotic stress experiments indicate that the binding of H10 is accompanied by the release of approximately 35 water molecules.We estimate from our naked DNA titration results that the binding of the H10 to the nucleosome places the H10 protein in close contact with approximately 41 DNA bp. The breakdown is that the H10 carboxyl terminus interacts with 28 bp of linker DNA on one side of the nucleosome, the H10 globular domain binds directly to 7 bp of core DNA, and shields another 6 linker DNA bases, 3 bp on either side of the nucleosome where the linker DNA exits the nucleosome core.

Approach to the Investigation of Nucleosome Structure by Using the Highly Emissive Nucleobase thdG–tC FRET Pair

“…Although fluorescence of DNA and nucleosome with acceptor (A 41st or A 27th) is similar (Figure S12, Supporting Information), nucleosome containing donor (D 40th) increased fluorescence intensity 1.89‐fold compared with duplex DNA. We estimated R DA based on a minimized structure using the software BIOVIA discovery studio version 17.1 . After minimization of th dG and tC‐containing nucleosome structure, the R DA of nucleosome with D 40th+A 41st (47.2±0.06 Å) and D 40th+A 27th (26.5±0.05 Å) was observed (Figure S9, Supporting Information).…”