Periodic binding of individual core histones to DNA: inadvertent purification of the core histone H2B as a putative enhancer-binding factor.
By using a DNase I footprinting assay, we have purified a factor by DNA affinity chromatography that binds to the minimal enhancer region of the Drosophila knirps gene and subsequently identified the protein as the core histone H2B. This inadvertent purification of a core histone as a putative sequence-specific DNA binding protein was due to a previously unknown property of H2B to interact with DNA in a periodic manner. Moreover, we found that each of the individual core histones, but not histone H1 or high mobility group protein 1, bound to the knirps enhancer to give a repetitive DNase I footprint pattern with a periodicity of about 10 base pairs, which is approximately one turn of the DNA helix. In addition, preparations containing the core histones H2A-H2B or H3-H4 yielded identical periodic DNase I footprint patterns on several different promoter and enhancer regions. These findings suggest that there are periodic, homotypic interactions between DNA-bound core histones that result from an alteration of the overall DNA structure such as the curvature rather than a specific sequence. We have also shown that histones H2A-H2B can repress initiation of transcription by RNA polymerase II. The phenomena described here may reflect histone-DNA interactions in non-nucleosomal stretches of chromatin and could be involved in some aspects of either rotational or translational positioning of nucleosomes. Furthermore, these findings indicate that a repeated 10 bp DNase I ladder, which has previously been considered to be a property of an intact nucleosome, can also be generated with subnucleosomal components. It will thus be necessary to reevaluate the criteria applied to the analysis of nucleosomes both in vivo and in vitro.
Kerrigan
L A
LA
Department of Biology, University of California, San Diego, La Jolla 92093.
Kadonaga
J T
JT
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002843:Chromatin / Q000502:physiology / Q000648:ultrastructure; D002846:Chromatography, Affinity; D004247:DNA / Q000378:metabolism*; D003850:Deoxyribonuclease I; D004330:Drosophila / Q000196:embryology / Q000235:genetics; D004625:Embryo, Nonmammalian; D004742:Enhancer Elements, Genetic; D006657:Histones / Q000302:isolation & purification* / Q000378:metabolism*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D012319:RNA Polymerase II / Q000378:metabolism; D013379:Substrate Specificity; D014158:Transcription, Genetic
D002843:Chromatin; D006657:Histones; D004247:DNA; D012319:RNA Polymerase II; D003850:Deoxyribonuclease I