Histone H5 can correctly align randomly arranged nucleosomes in a defined in vitro system.

In eukaryotic cells, DNA is packed into regularly spaced chromatin subunits called nucleosomes. The average distance between nucleosomes (the repeat length) varies in a tissue- and species-specific manner, with values ranging from about 160 to 240 DNA base pairs (bp). Thus, it has been recognized that the repeat length could be one of the factors underlying selective gene expression. In cells growing in culture, the characteristic repeat length for that type of cell seems to arise from an immature chromatin structure in which nucleosomes are initially irregularly spaced or are arranged in small closely packed clusters. At present no in vitro system has been described which is capable of reconstituting the mature physiological nucleosome spacing from purified chromatin components. Moreover, neither the factors necessary for spacing nor the reaction mechanism are known. We describe here an in vitro system that can restore the native subunit spacing in rearranged chromatin samples which have irregularly spaced nucleosomes similar to the situation apparent in newly replicated chromatin.