Molecular dynamics of DNA and nucleosomes in solution studied by fast-scanning atomic force microscopy.
Nucleosome is a fundamental structural unit of chromatin, and the exposure from or occlusion into chromatin of genomic DNA is closely related to the regulation of gene expression. In this study, we analyzed the molecular dynamics of poly-nucleosomal arrays in solution by fast-scanning atomic force microscopy (AFM) to obtain a visual glimpse of nucleosome dynamics on chromatin fiber at single molecule level. The influence of the high-speed scanning probe on nucleosome dynamics can be neglected since bending elastic energy of DNA molecule showed similar probability distributions at different scan rates. In the sequential images of poly-nucleosomal arrays, the sliding of the nucleosome core particle and the dissociation of histone particle were visualized. The sliding showed limited fluctuation within approximately 50nm along the DNA strand. The histone dissociation occurs by at least two distinct ways: a dissociation of histone octamer or sequential dissociations of tetramers. These observations help us to develop the molecular mechanisms of nucleosome dynamics and also demonstrate the ability of fast-scanning AFM for the analysis of dynamic protein-DNA interaction in sub-seconds time scale.
Suzuki
Yuki
Y
Laboratory of Plasma Membrane and Nuclear Signaling, Graduate School of Biostudies, Kyoto University, Sakyo-ku, Kyoto, Japan. ysuzuki79.m07@lif.kyoto-u.ac.jp
Higuchi
Yuji
Y
Hizume
Kohji
K
Yokokawa
Masatoshi
M
Yoshimura
Shige H
SH
Yoshikawa
Kenichi
K
Takeyasu
Kunio
K
D002843:Chromatin / Q000737:chemistry / Q000378:metabolism; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D006367:HeLa Cells; D006657:Histones / Q000378:metabolism; D006801:Humans; D018625:Microscopy, Atomic Force / Q000295:instrumentation / Q000379:methods*; D009690:Nucleic Acid Conformation; D009707:Nucleosomes / Q000737:chemistry / Q000378:metabolism*; D012996:Solutions / Q000378:metabolism*; D013816:Thermodynamics
D002843:Chromatin; D006657:Histones; D009707:Nucleosomes; D012996:Solutions; D004247:DNA