Nicking-closing enzyme assembles nucleosome-like structures in vitro.
The four core histones (H2A, H2B, H3, and H4) and DNA were assembled into nucleosome-like particles at physiological ionic strengths either by an extract of chromatin rich in nicking-closing activity or by the purified nicking-closing enzyme itself. When histone-DNA complexes were assembled in vitro from relaxed circular DNA, nearly physiological numbers of superhelical turns were induced in the DNA molecule. Electron microscopy of the complexes assembled by the chromatin extract revealed a beaded structure and a reduction of the contour length compared to free DNA. Micrococcal nuclease digestion of the histone-DNA complexes yielded 145-base-pair DNA fragments typical of nucleosome core particles and shorter subnucleosomal DNA fragments of discrete length.
D000818:Animals; D003086:Bacteriocin Plasmids; D002843:Chromatin / Q000648:ultrastructure; D004264:DNA Topoisomerases, Type I / Q000378:metabolism*; D004269:DNA, Bacterial / Q000378:metabolism; D004270:DNA, Circular / Q000378:metabolism; D004278:DNA, Superhelical / Q000378:metabolism; D004331:Drosophila melanogaster; D006657:Histones / Q000378:metabolism; D008836:Micrococcal Nuclease / Q000378:metabolism; D009707:Nucleosomes / Q000378:metabolism* / Q000648:ultrastructure; D051381:Rats
D002843:Chromatin; D004269:DNA, Bacterial; D004270:DNA, Circular; D004278:DNA, Superhelical; D006657:Histones; D009707:Nucleosomes; D008836:Micrococcal Nuclease; D004264:DNA Topoisomerases, Type I