Physicochemical studies of the folding of the 100 A nucleosome filament into the 300 A filament. Cation dependence.
The cation-induced refolding of the 100 A nucleosome filament into the 300 A filament has been studied over a wide range of concentrations of Na+, Mg2+, Co(NH3)3+6 and other cations. X-ray diffraction, electron microscopy and analytical ultracentrifugation have been used to determine the conditions under which the 300 A filament is formed. It is shown that cations induce chromatin refolding by acting as general DNA counterions. The concentration of any cation required to induce refolding is greatly dependent on the valence of that cation. Na+ (and, presumably, other monovalent cations) has dual effects: at high concentrations (greater than 45 to 65 mM) it stabilizes the 300 A filament state of chromatin; however, at low concentrations (less than approximately equal to 45 mM), when cations of higher valence are present and stabilizing the 300 A filament state, Na+ has the opposite effect, competing with the higher-valence cation for binding to the chromatin and destabilizing the 300 A filament state. It is shown that further addition of cations to chromatin in the 300 A filament state causes a further folding of the chromatin in which the sedimentation coefficient increases and the X-ray diffraction bands resulting from nucleosomal packing sharpen. This may reflect subtle structural changes within the 300 A filament, or it may reflect a shift in equilibrium constant for chromatin fluctuating between the 100 A and 300 A filament states. It is also shown that, with continued addition of cation, the 300 A filaments precipitate before any "endpoint" is reached in this further folding. The tendency of 300 A filaments to aggregate in vitro appears to be a built-in property, and may reflect the packing of 300 A filaments within metaphase chromosomes in vivo.
D000818:Animals; D002843:Chromatin / Q000648:ultrastructure*; D004247:DNA; D008274:Magnesium; D008854:Microscopy, Electron; D009690:Nucleic Acid Conformation; D009707:Nucleosomes / Q000032:analysis*; D012964:Sodium; D014961:X-Ray Diffraction
D002843:Chromatin; D009707:Nucleosomes; D004247:DNA; D012964:Sodium; D008274:Magnesium