Abstract
We have formed complexes of relaxed closed circular Col E1 DNA with various combinations of histones, and examined the effects of treating the complexes with nicking-closing enzyme. Germond et al (1) have shown that when a mixture of the four core histones of the nucleosome (HIA, H2B, H3 and H4) is used in such an experiment, the subsequently isolated DNA is supercoiled. We find that the arginine-rich histone pair, H3 and H4, is sufficient to induce the supercoiling observed in this experiment. Both H3 and H4 are required, and in the absence of either, no other histones are effective. H3 and and H4 are as efficient, per unit weight, as a mixture of the four histones in inducing supercoils. We also show that there is a large difference between the DNA bending energy needed to form a nucleosome and that needed to form one turn of normal superhelical DNA. These two processes are energetically quite distinct and probably separable. We estimate the free energy of interaction between DNA-bound histone pairs, and find that one or two such interactions would generate enough energy to fold the DNA into a nucleosome.
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