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Nucleic Acids Research, 1978, Vol. 5, No. 10 3603-3618
© 1978

Articles

Physical properties of inner histone-DNA complexes

P.N. Bryan*, E.B. Wright, M.H. Hsie, A.L. Olins and D.E. Olins

University of Tennessee-Oak Ridge Graduate School of Biomedical Sciences, and Biology Division, Oak Ridge National Laboratory Oak Ridge, TN 37830, USA

*To whom correspondence should be addressed

Received June 30, 1978. Chicken erythrocyte inner histone tetramer has been complexed with several natural and synthetic DNA duplexes by salt-gradient dialysis at various protein/DNA ratios. The resulting complexes, in low-ionic-strength buffer, have been examined by electron microscopy, circular dichroism, and thermal denaturation. Electron microscopy reveals nucleosomes ({varepsilon} bodies) randomly arranged along DNA fibers, including poly(dA-dT).poly(dA-dT), poly(dI-dC.poly(dI-dC), but not poly(dA).poly(dT). Circular dichroism studies showed prominent histone {alpha}-helix and "suppression" of nucleic acid ellipticity ({lambda}>240 nm). Thermal denaturation experiments revealed Tm behavior comparable to that of H1- (or H5-) depleted chromatin. Tm III and Tm IV increased linearly with G+C% (natural DNAs), but were virtually independent of the histone/DNA ratio; therefore, the melting of nucleosomes along a DNA chain is insensitive to adjacent "spacer" DNA lengths. This suggests that Tm III and Tm IV arise from the melting of different domains of DNA associated with the core {varepsilon} body.


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