ChromEMT: Visualizing 3D chromatin structure and compaction in interphase and mitotic cells
Year: 2017
Type: article
Abstract: The chromatin structure of DNA determines genome compaction and activity in the nucleus. On the basis of in vitro structures and electron microscopy (EM) studies, the hierarchical model is that 11-nanometer DNA-nucleosome polymers fold into 30- and subsequently into 120- and 300- to 700-nanometer fibers and mitotic chromosomes. To visualize chromatin in situ, we identified a fluorescent dye that stains DNA with an osmiophilic polymer and selectively enhances its contrast in EM. Using ChromEMT (ChromEM tomography), we reveal the ultrastructure and three-dimensional (3D) organization of individual chromatin polymers, megabase domains, and mitotic chromosomes. We show that chromatin is a disordered 5- to 24-nanometer-diameter curvilinear chain that is packed together at different 3D concentration distributions in interphase and mitosis. Chromatin chains have many different particle arrangements and bend at various lengths to achieve structural compaction and high packing densities.
Source: Science
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
Funders: Howard Hughes Medical Institute, W. M. Keck Foundation, Leona M. and Harry B. Helmsley Charitable Trust, Louis and Harold Price Foundation, National Institutes of Health, NIH Office of the Director, National Cancer Institute, National Institute of General Medical Sciences, National Institute of Biomedical Imaging and Bioengineering
Awards: G1599729373, G187995827, G2317269023, G2919467636, IVEM and Image Analysis Resource, G3337127418, G3377789135, G3679753445, G4084676586, San Diego Center for Systems Biology: From Maps to Models, G4864671638, FEI TITAN HALO 300kV INTERMEDIATE VOLTAGE ELECTRON MICROSCOPE, G5457882003, G680976502, Stochastic gene expression in differentiation and development, G7814828079, G8415296469, Cancer Center Support Grant (CCSG)
OpenAlex ID: W2739969873