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Free keywords:
NUCLEOSOME CORE PARTICLES; X-RAY-DIFFRACTION; MITOTIC CHROMOSOMES;
COMPACT; FIBERS; DNA; ORGANIZATION; MICROSCOPY; REVEALS; ORDERBiochemistry & Molecular Biology; Cell Biology; chromatin higher-order structure; cryo-electron tomography; DNA
packaging; metaphase chromosome structure; small-angle X-ray scattering;
Abstract:
Cryo-electron tomography and small-angle X-ray scattering were used to investigate the chromatin folding in metaphase chromosomes. The tomographic 3D reconstructions show that frozen-hydrated chromatin emanated from chromosomes is planar and forms multilayered plates. The layer thickness was measured accounting for the contrast transfer function fringes at the plate edges, yielding a width of similar to 7.5 nm, which is compatible with the dimensions of a monolayer of nucleosomes slightly tilted with respect to the layer surface. Individual nucleosomes are visible decorating distorted plates, but typical plates are very dense and nucleosomes are not identifiable as individual units, indicating that they are tightly packed. Two layers in contact are similar to 13 nm thick, which is thinner than the sum of two independent layers, suggesting that nucleosomes in the layers interdigitate. X-ray scattering of whole chromosomes shows a main scattering peak at similar to 6 nm, which can be correlated with the distance between layers and between interdigitating nucleosomes interacting through their faces. These observations support a model where compact chromosomes are composed of many chromatin layers stacked along the chromosome axis.
