Studies on histone oligomers. V. Reconstitution of chromatin from purified DNA and acid-extracted histones.
DNA-histone complexes were reconstituted from DNA and acid-extracted core histones and the products were characterized by micrococcal nuclease digestion to examine whether proper nucleosome structure had been reconstituted. No nucleosome structure was produced starting from the mixture of acid-extracted histones and purified DNA in 2 M NaCl-5 M urea, while the reassociation of chromatin by the same procedures was successful. This was due to the inappropriate conformation of acid-extracted histones, which was preserved in 2 M NaCl even in the presence of 5 M urea. If acid-extracted histones were reannealed from the completely denatured state, such as in 5 M urea, 6 M guanidine hydrochloride or 0.6 M NaCl-5 M urea, reconstitution of nucleosome structure was always successful.
D000818:Animals; D002645:Chickens; D002843:Chromatin / Q000378:metabolism / Q000648:ultrastructure*; D004247:DNA / Q000378:metabolism*; D004912:Erythrocytes / Q000378:metabolism; D006657:Histones / Q000378:metabolism*; D007700:Kinetics; D046911:Macromolecular Substances; D008836:Micrococcal Nuclease; D009691:Nucleic Acid Denaturation; D009707:Nucleosomes / Q000378:metabolism*; D011489:Protein Denaturation
D002843:Chromatin; D006657:Histones; D046911:Macromolecular Substances; D009707:Nucleosomes; D004247:DNA; D008836:Micrococcal Nuclease