Studies on the thermal denaturation of histone-H1-depleted chromatin.
The multiphasic thermal denaturation profile of histone-H1-depleted chromatin was studied by using a nucleoprotein preparation which lacked the first high temperature transition at about 72 degrees C. Such a preparation was obtained by heating at 72 degrees C H1-depleted chromatin, the DNA of which was cross-linked with psoralen to ensure a complete renaturation of DNA upon cooling. When this nucleoprotein was redenatured, its melting profile was found to be significantly altered: only one high temperature peak centered at about 82 degrees C was observed in addition to the low temperature transition at about 53 degrees C. The kinetics of digestion of this material with micrococcal nuclease showed a limit digest equal to that found for the 'native' H1-depleted chromatin but the rate of hydrolysis was higher. The monomer particles prepared from this nucleoprotein were found similar to the 'native' monosomes in respect to protein:DNA ratio and size of DNA but showed an altered melting profile: the premelt area was broader, bigger, and centered at lower temperature; the main peak was reduced in size with no change in its melting temperature. On the basis of these data, the following conclusions were drawn: (a) the last two thermal transitions in H1-depleted chromatin most likely reflect the presence within each nucleosome of two regions with different stability of DNA; (b) DNA involved in the first high thermal transition of H1-depleted chromatin belongs to nuclease-accessible DNA, and (c) the main peak in the biphasic melting profile of the monomer particles reflects the denaturation of DNA regions which in the polymer nucleoprotein are involved in the two high temperature transitions.
D000818:Animals; D002467:Cell Nucleus / Q000648:ultrastructure; D002843:Chromatin / Q000648:ultrastructure*; D004247:DNA / Q000032:analysis*; D006657:Histones; D007700:Kinetics; D008099:Liver / Q000032:analysis; D008836:Micrococcal Nuclease / Q000378:metabolism; D009691:Nucleic Acid Denaturation; D009707:Nucleosomes / Q000648:ultrastructure; D011489:Protein Denaturation; D051381:Rats; D013696:Temperature
D002843:Chromatin; D006657:Histones; D009707:Nucleosomes; D004247:DNA; D008836:Micrococcal Nuclease