DNA sequence directs placement of histone cores on restriction fragments during nucleosome formation

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Abstract
Restriction fragments, 203 and 144 base pairs in length, bearing the Escherichia coli lac control region were reconstituted with the core histones from calf thymus to form nucleosomes. By several criteria the reconstituted nucleosomes are similar to native nucleosomes obtained by micrococcal nuclease digestion of calf thymus nuclei. Sensitive nuclease digestion studies reveal subtle and important differences between native monosomes and the lac reconstitutes. Each reconstitute consists mainly of nucleosomes containing histone cores placed nonrandomly with respect to the DNA sequence. The shorter reconstitute forms asymmetric nucleosomes as evidenced by the DNase I digestion pattern. Exonuclease III digestion followed by 5''-end analysis of the larger reconstitute suggest that, of the many possible arrangements of histone core with DNA sequence, only 2 are highly favored.