Co-operative binding of the globular domain of histone H5 to DNA.
The globular domain of histone H5 (GH5) was prepared by trypsin digestion of H5 that was extracted from chicken erythrocyte nuclei with NaCl. Electron microscopy, sucrose gradient centrifugation, native agarose gel electrophoresis and equilibrium density gradient ultracentrifugation show that GH5 binds co-operatively to double-stranded DNA. The electron microscopic images suggest that the GH5-DNA complexes are very similar in structure to co-operative complexes of intact histone H1 (or its variants) with double-stranded DNA, studied previously, which have been proposed to consist of two parallel DNA double helices sandwiching a polymer of the protein. For complexes with GH5 or with intact H1, naked DNA co-sediments with the protein-DNA complexes through sucrose gradients, and DNA also appears to protrude from the ends and sides of the complexes; measurements of the protein-DNA stoichiometry in fractionated samples may not reflect the stoichiometry in the complexes. An estimate of the stoichiometry obtained from the buoyant density of fixed GH5-DNA complexes in CsCl suggests that sufficient GH5 is present in the complexes for the GH5s to be in direct contact, as required by a simple molecular mechanism for the co-operative binding. Chemical crosslinking demonstrates that GH5s are in close proximity in the complexes. In the absence of DNA, GH5-GH5 interactions are weak or non-existent.
Draves
P H
PH
Department of Chemistry, University of Illinois, Urbana-Champaign 61801.
Lowary
P T
PT
Widom
J
J
D000818:Animals; D001665:Binding Sites; D002467:Cell Nucleus / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D002645:Chickens; D002843:Chromatin / Q000378:metabolism; D004247:DNA / Q000302:isolation & purification / Q000378:metabolism* / Q000648:ultrastructure; D004912:Erythrocytes / Q000378:metabolism; D006657:Histones / Q000302:isolation & purification / Q000378:metabolism* / Q000648:ultrastructure; D007700:Kinetics; D008027:Light; D008854:Microscopy, Electron; D012542:Scattering, Radiation
D002843:Chromatin; D006657:Histones; D004247:DNA