Histone-H1-dependent chromatin superstructures and the suppression of gene activity

I have identified a chromatin particle containing DNA as large as 20-40 kb that migrates as a discrete entity on agarose gels. With increasing nuclease digestion, the particle becomes cleaved in the linker regions between nucleosomes, but remains intact, probably held together by the outer histones, H1 and H5. By hybridization analysis, inactive genes are found in these particles. Active genes (and their flanking sequences) are also found in particles containing H1 and H5, but in contrast to inactive supranucleosome particles, active polynucleosome particles are not held together after cleavage of linker DNA. This suggests that H1 cross-links adjacent nucleosomes in inactive regions and that H1 is bound differently in expressed regions. The results raise the possibility that the marked degree of suppression of repressed, tissue-specific genes may be determined, in part, by their assembly into these inactive supranucleosome structures.     

Structure of transcriptionally-active chromatin subunits.

Rat liver chromatin is organized into regions of DNA which differ in degree of susceptibility to attack by the endonucleases DNase I and DNase II. The most nuclease-sensitive portion of chromatin DNA is enriched in transcribed sequences. This fraction may be separated from the bulk of chromatin by virtue of its solubility in solutions containing 2 mM MgCl2. Both transcribed and nontranscribed regions of chromatin are organized into repeating units of DNA and histone, which appear as 100 A beads in the electron microscope. The length of DNA in the repeat unit is the same for these two classes of chromatin (198 +/- 6 base pairs in rat liver); however, the subunits of active, Mg++-soluble chromatin differ from the nucleosomes of inactive regions of chromatin in several respects. Active subunits are enriched in nascent RNA and nonhistone protein and exhibit higher sedimentation values than the corresponding subunits of inactive chromatin.