Thermal denaturation and template activities of reconstituted DNA-histone complexes.

Reconstituted complexes of DNA with histone were prepared by salt-and-urea step gradient dialysis. The DNA was complexed with histone H1, with the combination of the other four histones H2A, H2B, H3 and H4, and with whole histones. These DNA-histone complexes were purified by Bio-Gel column chromatography, and the weight ratio of histone-to-DNA was determined in each complex. The thermal denaturation profile and nuclease digestion pattern of DNA-histone H2A, H2B, H3 and H4 complex were compatible with those of the polynucleosome structure of chromatin. The template activities for transcription were compared in these DNA-histone complexes by separately measuring initiation reaction and chain elongation. The binding of histone H1 to DNA strongly inhibited the initiation, while the binding of the combination of the other four histones to DNA partially inhibited the initiation and chain elongation. The binding characteristics are discussed with regard to the role of histone H1 and the other four histones in chromatin structure and template activity.