Contribution of histones H2A and H2B to the folding of nucleosomal DNA

Chemicals and CAS Registry Numbers deoxyribonuclease I, 9003-98-9; dimethylmaleic acid anhydride, 766-39-2; DNA, 9007-49-2; histone, 9062-68-4; lysine, 56-87-1, 6899-06-5, 70-54-2; maleic anhydride, 108-31-6; 2,3-dimethylmaleic anhydride, 766-39-2; Deoxyribonuclease I, EC 3.1.21.1; DNA, 9007-49-2; Histones; Lysine, 56-87-1; Maleic Anhydrides; Nucleosomes

We have studied the structural properties of nucleosomal particles deficient in histones H2A and H2B produced by modification of histone amino groups with dimethylmaleic anhydride [Jordano, J., Montero, F., & Palacián, E. (1984) Biochemistry (preceding paper in this issue)]. Digestion with DNase I of residual particles containing only 15% of the original H2A·H2B complement produces only discrete DNA fragments no longer than 70 nucleotides. As compared with the original nucleosomes, thermal denaturation of the residual particles shows a decrease from 140 to about 90 in the number of nucleotide base pairs per particle that melt at the highest temperature transition as well as a drop in the temperature of this transition. Circular dichroism spectra of the residual particles give ellipticity values around 275 nm, much higher than those corresponding to the control nucleosomes, which appears to indicate a loss in the compact DNA tertiary structure. When regeneration of the modified amino groups of the residual particles takes place in the presence of the complementary fraction containing histones H2A and H2B, but not in its absence, nucleosomal particles with the structural properties of the original nucleosomes are reconstituted. Therefore, the structural change observed in the residual particles can be assigned to the lack of histones H2A and H2B and not to the modified amino groups of the histones present in the residual particles. The results are consistent with the stabilization by histones H2A and H2B of a DNA length of 50-70 base pairs per nucleosome.

This work was supported in part by the Fondo de Investigaciones Sanitarias and the Comisión Asesora de Investigación Cientifica y Técnica (Spain).

Peer reviewed