Presence of nucleosomes within irregularly cleaved fragments of newly replicated chromatin

In previous reports (Annunziato et al., J. Biol. Chem., 256:11880-11886 [1981]; Annunziato and Seale, Biochemistry 21:5431-5438 [1982]) we have described two classes of newly replicated chromatin which differ in structure, solubility properties, and requirements for maturation. One class is nucleosomal, soluble at low to intermediate ionic strengths, and acquires mature nucleosomal composition and normal repeat length in the absence of concurrent protein synthesis. In contrast, the other class is cleaved irregularly by MNase (appearing as a smear in DNA gels), is insoluble at moderate ionic strengths, requires protein synthesis to gain normal subunit structure, and comprises approximately 60% of total new chromatin DNA after mild nuclease digestion. It is now demonstrated that this heterogeneous component (produced by the action of either MNase or Hae III on chromatin replicated in cycloheximide) yields nucleosomes when redigested with MNase. The presence of nucleosomes within heterogeneous chromatin fragments suggests that nucleosomal and non-nucleosomal regions may be juxtaposed during chromatin replication. These findings are discussed with respect to current models of nucleosome segregation.