The integrity of the histone-DNA complex in chromatin fibres is not necessary for the maintenance of the shape of mitotic chromosomes.
In this study we addressed the question of whether scaffold structures produced from purified mitotic chromosomes are an artefact of dehistonization, and whether the integrity of the chromatin fibres is necessary for the maintenance of the well-known shape of mitotic chromosomes. Purified mitotic chromosomes from Friend erythroleukemia cells were treated either with increasing NaCl concentrations up to 500 mM, or with 6 M urea in the presence or absence of 10 mM 2-mercaptoethanol. The main criterion for the intactness of the overall chromosome shape as seen by electron microscopy was the characteristic X- or U-like appearance with clearly discernable chromatid axes. Histone H1 is known to be essential for the integrity of chromatin fibres. Its removal in sucrose gradients containing 500 mM NaCl did not lead to loss of the overall chromosome shape. However, treatment of chromosomes in sucrose gradients containing 10 mM 2-mercaptoethanol and 6 M urea led to loss of the structure probably due to dissociation (or denaturation) of shape-determining (scaffolding) components. Under these conditions most of the histones remained bound to the chromosomes, and the fibres in this chromatin material, after removal of excess urea and 2-mercaptoethanol, still showed condensation of the nucleosome filaments into the characteristic fibre structures upon increasing ionic strength. Our observations are compatible with the model that specific non-histone components, independently of histone-DNA interactions, organize or stabilize the structure of metaphase chromosomes.
Homberger
H
H
Institut für Zellbiologie, Eidgenössische Technische Hochschule Zürich (ETHZ), Switzerland.
Koller
T
T
D000818:Animals; D002843:Chromatin / Q000378:metabolism*; D002875:Chromosomes / Q000378:metabolism* / Q000648:ultrastructure; D004247:DNA / Q000378:metabolism*; D005622:Friend murine leukemia virus; D006657:Histones / Q000378:metabolism*; D004915:Leukemia, Erythroblastic, Acute / Q000378:metabolism / Q000473:pathology; D008623:Mercaptoethanol; D008854:Microscopy, Electron; D008938:Mitosis; D012965:Sodium Chloride; D014407:Tumor Cells, Cultured / Q000378:metabolism / Q000648:ultrastructure; D014508:Urea
D002843:Chromatin; D006657:Histones; D012965:Sodium Chloride; D008623:Mercaptoethanol; D014508:Urea; D004247:DNA