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Nucleic Acids Research, 1977, Vol. 4, No. 5 1159-1182
© 1977

Articles

Supercoiling energy and nucleosome formation: the role of the arginine-rich histone kernel*,{dagger}

R.Daniel Camerini-Otero and Gary Felsenfeld

Laboratory of Molecular Biology, National Institute of Arthritis, Metabolism and Digestive Diseases, National Institutes of Health Bethesda, MD 20014, USA

Received February 4, 1977.

We have formed complexes of relaxed closed circular Col El 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 (H2A, 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 HA 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.

*Dedicated to the memory of Jerome Vinograd, a good friend and teacher.

{dagger}A preliminary report of some of these results was presented at a Symposium on Chromatin Structure and Function at the First International Congress on Cell Biology, Boston, Massachusetts, September 6, 1976. Felsenfeld, C., Camerini-Otero, R.D. and Sollner-Webb, B. (1977) in "International Cell Biology 1976-1977".Rockefeller University Press, B. Brinkley and K. Porter (eds.), in press.


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