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J. Biochem, 1984, Vol. 96, No. 4 1071-1078
© 1984 Japanese Biochemical Society

research-article

Reconstitution of Compact Polynucleosomes and Comparison of the Functions of Histones H1 and H5

Koichiro TAKASHIMA*,1, Seiichi KAWASHIMA**,2 and Kazutomo IMAHORI**

*Department of Biochemistry, Faculty of Medicine, The University of Tokyo Bunkyo-ku Tokyo 113
**Department of Biochemistry, Tokyo Metropolitan Institute of Gerontology Sakae-cho, Itabashi-ku, Tokyo 173

2To whom all correspondence should be addressed. Abbreviations; SDS, Sodium dodecyl sulfate; EGTA, ethyleneglycol-bis-ß-aminoethyl ether)-N, N, N', N'-tetra-acetic acid; EDTA, ethylenediamine tetracetic acid.

Polynucleosomes with a definite length (about 4, 500 base pairs) were prepared from chicken erythrocyte nuclei without depleting magnesium ions from the medium. The polynucleosomes in the presence of Mg2+ ions as well as monovalent salts were more compact than those with monovalent salts alone. We minimized the occurrence of nicks in the DNA of nucleosome fiber during the preparation. When histones H1 and H5 were completely removed from polynucleosomes, linker histonedepleted polynucleosomes sedimented slower than the original ones. When isolated histone H1 or H5 was reassembled with linker histone-depleted polynucleosomes, no significant difference was observed among the reconstituted polynucleosomes with histone H1, the reconstituted polynucleosomes with histone H1 and H5 are similar in their effects on higher order structure of polynucleosomes, as far as can be judged from such characteristics as sedimentation velocity, linker histone content, and the patterns of nuclease digestion.

1Department of Physiology, Faculty of Medicine, Iwate Medical University, Uchimaru, Morioka, Iwate 020


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