Control of RNA polymerase binding to chromatin by variations in linker histone composition.
We have measured the frequency of initiation sites in chromatin for RNA polymerase in vitro as a function of the composition of linker histones (H1 and its analogues). In linker histone-depleted chromatin, RNA chain initiation appears to be restricted to the exposed linker DNA. On titration with purified linker histones, initiation is further restricted to an extent determined by the amount and type of linker histone, and the source of depleted chromatin. The extent of repression is correlated with the capacity of linker histones to induce the formation of higher-order structure in the complex. The results suggest that the effects of linker histones are mediated through the higher-order structure of chromatin, which prevents access of polymerase to the linker DNA. Accordingly, we find that structures imposed by the linker histones after polymerase binding are not inhibitory. Microscopy reveals that the higher-order structure in partially condensed chromatin is discontinuous, with solenoidal units spaced by sections of unravelled nucleosomes. Since salt stimulation of linker histone exchange does not result in derepression of linkers in our assay, we conclude that the distribution of higher-order units in chromatin is static and that the linker histones exchange between high-affinity sites in established units. We have previously shown that the globin gene is selectively unfolded in tissues that express the gene. The present results suggest that the transcriptional activity of specific genes is maintained by differential linker histone binding within chromatin.
D000818:Animals; D001667:Binding, Competitive; D002417:Cattle; D002843:Chromatin / Q000378:metabolism* / Q000648:ultrastructure; D004247:DNA; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology; D006657:Histones / Q000378:metabolism*; D008854:Microscopy, Electron; D012319:RNA Polymerase II / Q000378:metabolism; D013698:Templates, Genetic; D014461:Ultracentrifugation
D002843:Chromatin; D006657:Histones; D004247:DNA; D012319:RNA Polymerase II; D012321:DNA-Directed RNA Polymerases