Atomic force microscopy sees nucleosome positioning and histone H1-induced compaction in reconstituted chromatin.
We addressed the question of how nuclear histones and DNA interact and form a nucleosome structure by applying atomic force microscopy to an in vitro reconstituted chromatin system. The molecular images obtained by atomic force microscopy demonstrated that oligonucleosomes reconstituted with purified core histones and DNA yielded a 'beads on a string' structure with each nucleosome trapping 158 +/- 27 bp DNA. When dinucleosomes were assembled on a DNA fragment containing two tandem repeats of the positioning sequence of the Xenopus 5S RNA gene, two nucleosomes were located around each positioning sequence. The spacing of the nucleosomes fluctuated in the absence of salt and the nucleosomes were stabilized around the range of the positioning signals in the presence of 50 mM NaCl. An addition of histone H1 to the system resulted in a tight compaction of the dinucleosomal structure.
Sato
M H
MH
Department of Natural Environmental Sciences, Faculty of Integrated Human Studies, Kyoto University, Japan.
Ura
K
K
Hohmura
K I
KI
Tokumasu
F
F
Yoshimura
S H
SH
Hanaoka
F
F
Takeyasu
K
K
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism / Q000648:ultrastructure; D002843:Chromatin / Q000378:metabolism / Q000648:ultrastructure*; D004247:DNA / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D004275:DNA, Ribosomal / Q000235:genetics; D006367:HeLa Cells; D006657:Histones / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D006801:Humans; D018625:Microscopy, Atomic Force / Q000379:methods; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D009707:Nucleosomes / Q000378:metabolism / Q000648:ultrastructure*; D011487:Protein Conformation; D012341:RNA, Ribosomal, 5S / Q000235:genetics; D013698:Templates, Genetic; D014981:Xenopus
D002843:Chromatin; D004275:DNA, Ribosomal; D006657:Histones; D009707:Nucleosomes; D012341:RNA, Ribosomal, 5S; D004247:DNA