Irregular Folding of Human Genome DNA in the Cells

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How is 2-m-long genomic DNA organized into a mitotic chromosome or nucleus? The nucleosome fiber has long been assumed to be folded into a 30-nm chromatin fiber, and further helically folded larger fiber. However, when we observed frozen hydrated human mitotic cells using cryoelectron microscopy (cryo-EM), no higher-order structures including 30-nm chromatin fibers were found. To further investigate the bulk structure of mitotic chromosomes, we performed small-angle X-ray scattering (SAXS) at SPring-8. No structural feature larger than 11 nm was detected, even on a chromosome-diameter scale (~1 μm). We also found a similar scattering pattern in interphase nuclei of HeLa cells in the range up to ~275 nm. Our findings suggest a common structural feature in interphase and mitotic chromatins: compact and irregular folding of nucleosome fibers occurs without a 30-nm chromatin structure.