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Mol Cell. 2017 Jul 20;67(2):282-293.e7. doi: 10.1016/j.molcel.2017.06.018. Epub 2017 Jul 14.

Dynamic Organization of Chromatin Domains Revealed by Super-Resolution Live-Cell Imaging.

Author information

1
Biological Macromolecules Laboratory, Structural Biology Center, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan; Institute for Advanced Biosciences, Keio University, Fujisawa 252-8520, Japan.
2
Biological Macromolecules Laboratory, Structural Biology Center, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan; Department of Genetics, School of Life Science, Sokendai (Graduate University for Advanced Studies), Mishima, Shizuoka 411-8540, Japan.
3
Biological Macromolecules Laboratory, Structural Biology Center, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan.
4
Eugene Bell Center for Regenerative Biology and Tissue Engineering, Marine Biological Laboratory, Woods Hole, MA 02543, USA.
5
XFEL Utilization Division, Japan Synchrotron Radiation Research Institute (JASRI), Sayo-gun, Hyogo 679-5198, Japan.
6
Institute for Advanced Biosciences, Keio University, Fujisawa 252-8520, Japan.
7
Division of Molecular Cell Engineering, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan.
8
Department of Cell Biology, Erasmus MC, 3000 CA Rotterdam, the Netherlands.
9
Laboratory for Cell Polarity Regulation, Quantitative Biology Center, RIKEN, Suita, Osaka 565-0874, Japan.
10
The Institute of Scientific and Industrial Research, Osaka University, Ibaraki, Osaka 567-0047, Japan.
11
Biological Macromolecules Laboratory, Structural Biology Center, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan; Department of Genetics, School of Life Science, Sokendai (Graduate University for Advanced Studies), Mishima, Shizuoka 411-8540, Japan. Electronic address: kmaeshim@nig.ac.jp.

Abstract

The eukaryotic genome is organized within cells as chromatin. For proper information output, higher-order chromatin structures can be regulated dynamically. How such structures form and behave in various cellular processes remains unclear. Here, by combining super-resolution imaging (photoactivated localization microscopy [PALM]) and single-nucleosome tracking, we developed a nuclear imaging system to visualize the higher-order structures along with their dynamics in live mammalian cells. We demonstrated that nucleosomes form compact domains with a peak diameter of ∼160 nm and move coherently in live cells. The heterochromatin-rich regions showed more domains and less movement. With cell differentiation, the domains became more apparent, with reduced dynamics. Furthermore, various perturbation experiments indicated that they are organized by a combination of factors, including cohesin and nucleosome-nucleosome interactions. Notably, we observed the domains during mitosis, suggesting that they act as building blocks of chromosomes and may serve as information units throughout the cell cycle.

KEYWORDS:

PALM; chromatin; chromatin domain; chromatin dynamics; chromosome; single-nucleosome tracking

PMID:
28712725
DOI:
10.1016/j.molcel.2017.06.018
[Indexed for MEDLINE]
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