ABSTRACT
270-MHz proton magnetic resonance has been used to study the effect of phosphorylation of histone H1 in vitro on the structure of isolated H1 molecules and on the interaction of H1 with DNA. Phosphorylation at serine-105, which is located in the globular region of H1, was found to reduce the enthalpy of structure formation from 24 +/- 2 kcal mol-1 (100 +/- 8 kJ mol-1) to 13 +/- 2 kcal mol-1 (55 +/- 8 kJ mol-1). Phosphorylation at either or both of serine-37 and serine-105 was found to reduce the strength of binding of the histone to DNA considerably at some ionic strengths.
Subject(s)
Chromatin/ultrastructure , Histones , Lysine , Phosphoproteins , Animals , Cattle , DNA , Magnetic Resonance Spectroscopy , Nucleic Acid Conformation , Protein Conformation , Thermodynamics , Thymus GlandABSTRACT
The nuclear magnetic resonance (NMR) spectrum of chromatin at ionic strengths below about 0.5 M may be attributed solely to its histone H1 component. The effect of various ions and urea on the complex has been investigated using NMR and confirm that the contraction of the complex on increase of ionic strength is largely due to electrostatic interactions. A detailed study of the H1 - DNA complex has also been undertaken. The behaviour of H1 in the two cases is virtually identical, implying that in chromatin the H1 is complexed with the DNA rather than with the other histones. Microcalorimetric measurements reveal that the binding of H1 to DNA is athermic or involves a heat of reaction which is very small indeed.
Subject(s)
Chromatin/ultrastructure , DNA , Histones , Animals , Binding Sites , Calcium Chloride , Calorimetry , Cations, Divalent , Cations, Monovalent , Cattle , DNA/analysis , Histones/analysis , Lanthanum , Magnetic Resonance Spectroscopy , Nucleic Acid Conformation , Protein Binding , Protein Conformation , Thymus Gland , UreaABSTRACT
Conformational changes in histone H2A (ALK, F2A2, IIbl) as a function of ionic strength and pH have been followed using high resolution nuclear magnetic resonance (NMR), circular dichroism (CD), and infrared (ir). While change in pH from 3 to 7 (no added salt) causes little structural change, added salt induces the formation of both alpha helix (28 percent maximum) and intermolecular associates in the region of the molecule between 25 and 113. No beta structure was observed at high salt. By the use of different salts it was shown that the structural changes were due largely to nonspecific counterion screening by the added anion. Comparison of observed with simulated NMR spectra has led to the proposal that an ionic strength dependent equilibrium exists between largely unstructured coil molecules and fully structured and aggregated molecules. NMR spectra of H2A obtained in the presence of DNA showed that both the N- and C-terminal regions bind to DNA, i.e., not the portion of the chain that is involved in interhistone interactions.
Subject(s)
Histones , Amino Acids/analysis , Animals , Binding Sites , Cattle , Circular Dichroism , Histones/metabolism , Hydrogen-Ion Concentration , Macromolecular Substances , Magnetic Resonance Spectroscopy , Models, Molecular , Osmolar Concentration , Protein Binding , Protein Conformation , Sea Urchins , Sodium Chloride , Spectrophotometry, Infrared , Thymus GlandABSTRACT
Proton magnetic resonance, circular dichroism and other studies of whole and cleaved calf thymus histone H1 (formerly F1) reveal the presence of specific folded structures in the region approximately from residue 40--115. Ionic, hydrogen-bond and hydrophobic interactions all appear to contribute to the stability of the structure, which is predicted to contain alpha-helices in regions 42--55 and 58--75. No evidence was found for beta-structures, either inter or intramolecular, or for any structure formation outside the region 40--115. At 18 degrees C and a protein concentration of 2 mM the first-order exchange rate between random-coil and structured forms is slower than 80 s-1; at 40 degrees C the exchange rate is faster than 330 s-1.
