The interaction of histone H5 and its globular domain with core particles, depleted chromatosomes, polynucleosomes, and a DNA decamer.
Certain features of linker histone behavior were analyzed using a precipitation and a nitrocellulose filter binding assay. Chromatosomes, depleted of the linker histones, present one unique binding site to the globular domain of histone H5 (GH5) which involves the two 10-base pair DNA ends of the chromatosome. Additional binding to lower affinity sites is intrinsically different and results in aggregation as does all binding to core particles. These findings, as well as the binding study on a synthetic DNA decamer, lend support to earlier hypotheses of more than one DNA binding site on the globular domain. Our studies provide a deeper insight into the long standing question of H5/nucleosome stoichiometry. A salt dependence analysis of GH5 binding to H5-depleted chromatosomes indicates that GH5 displaces a number of ions similar to the total H1 linker histone, suggesting a delocalized binding of the carboxyl- and amino-terminal tails.
Segers
A
A
Instituut voor Molekulaire Biologie, Vrije Universiteit Brussel, Belgium.
Muyldermans
S
S
Wyns
L
L
D000818:Animals; D002645:Chickens; D002843:Chromatin / Q000737:chemistry / Q000648:ultrastructure*; D003852:Deoxyribonucleoproteins; D005374:Filtration; D006657:Histones / Q000378:metabolism*; D066298:In Vitro Techniques; D009707:Nucleosomes / Q000737:chemistry / Q000648:ultrastructure*; D009838:Oligodeoxyribonucleotides / Q000378:metabolism*; D009994:Osmolar Concentration; D010446:Peptide Fragments / Q000378:metabolism; D010969:Plastics; D012995:Solubility
D002843:Chromatin; D003852:Deoxyribonucleoproteins; D006657:Histones; D009707:Nucleosomes; D009838:Oligodeoxyribonucleotides; D010446:Peptide Fragments; D010969:Plastics