3D-CLEM Reveals that a Major Portion of Mitotic Chromosomes Is Not Chromatin

Booth, DG, Beckett, AJ ORCID: 0000-0001-8377-325X, Molina, O, Samejima, I, Masumoto, H, Kouprina, N, Larionov, V, Prior, IA ORCID: 0000-0002-4055-5161 and Earnshaw, WC
(2016) 3D-CLEM Reveals that a Major Portion of Mitotic Chromosomes Is Not Chromatin. Molecular cell, 64 (4). pp. 790-802.

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Recent studies have revealed the importance of Ki-67 and the chromosome periphery in chromosome structure and segregation, but little is known about this elusive chromosome compartment. Here we used correlative light and serial block-face scanning electron microscopy, which we term 3D-CLEM, to model the entire mitotic chromosome complement at ultra-structural resolution. Prophase chromosomes exhibit a highly irregular surface appearance with a volume smaller than metaphase chromosomes. This may be because of the absence of the periphery, which associates with chromosomes only after nucleolar disassembly later in prophase. Indeed, the nucleolar volume almost entirely accounts for the extra volume found in metaphase chromosomes. Analysis of wild-type and Ki-67-depleted chromosomes reveals that the periphery comprises 30%-47% of the entire chromosome volume and more than 33% of the protein mass of isolated mitotic chromosomes determined by quantitative proteomics. Thus, chromatin makes up a surprisingly small percentage of the total mass of metaphase chromosomes.

Item Type: Article
Uncontrolled Keywords: 3D-CLEM, chromosome periphery, Ki-67, electron microscopy, CLEM
Depositing User: Symplectic Admin
Date Deposited: 18 Nov 2016 11:54
Last Modified: 19 Jan 2023 07:25
DOI: 10.1016/j.molcel.2016.10.009
Open Access URL: http://www.sciencedirect.com/science/article/pii/S...
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URI: https://livrepository.liverpool.ac.uk/id/eprint/3004600