3D-CLEM Reveals that a Major Portion of Mitotic Chromosomes Is Not Chromatin.
3D-CLEM Reveals that a Major Portion of Mitotic Chromosomes Is Not Chromatin.
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DOI：
10.1016/j.molcel.2016.10.009
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发表时间：
2016-11-17
期刊：
影响因子：
16
通讯作者：
Earnshaw, William C.
中科院分区：
文献类型：
--
作者：
Booth, Daniel G.;Beckett, Alison J.;Molina, Oscar;Samejima, Itaru;Masumoto, Hiroshi;Kouprina, Natalay;Larionov, Vladimir;Prior, Ian A.;Earnshaw, William C.
近年来的研究揭示了Ki-67和染色体外周在染色体结构和分离中的重要性，但对这种难以捉摸的染色体隔室知之甚少。在这里，我们使用相关的光和连续块面扫描电子显微镜，我们称之为3D-CLEM，以超结构分辨率模拟整个有丝分裂染色体补体。前期染色体表面高度不规则，体积小于中期染色体。这可能是因为外周细胞的缺失，外周细胞只有在前期核仁解体后才与染色体结合。事实上，核仁体积几乎完全解释了中期染色体中发现的额外体积。对野生型和ki -67缺失染色体的分析表明，外围区占整个染色体体积的30%-47%，占分离有丝分裂染色体蛋白质量的33%以上。因此，染色质在中期染色体的总质量中所占的比例非常小。3D-CLEM结合了光学和连续块面扫描电子显微镜，所有46条人类染色体的完整结构已经确定，大部分有丝分裂染色体不是由染色质组成，光学和电子显微镜测定的染色体体积差异很大，Booth等人开发了一种称为3D-CLEM的显微镜方法。利用这种方法，他们发现有丝分裂染色体的很大一部分不是由染色质组成的，这挑战了传统的染色体观念。
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. 3D-CLEM combines light and serial block-face scanning electron microscopy The complete architecture of all 46 human chromosomes has been defined A large portion of mitotic chromosomes is not composed of chromatin Chromosome volumes determined by light and electron microscopy differ dramatically Booth et al. develop a microscopy method called 3D-CLEM. Using this approach, they find that a large portion of mitotic chromosomes is not composed of chromatin, challenging classical perceptions of chromosomes.
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DOI：
10.1038/nrg3454
发表时间：
2013-06
期刊：
Nature reviews. Genetics
影响因子：
--
作者：
通讯作者：
--
影响因子：
64.8
作者：
Cuylen S;Blaukopf C;Politi AZ;Müller-Reichert T;Neumann B;Poser I;Ellenberg J;Hyman AA;Gerlich DW
通讯作者：
Gerlich DW
DOI：
10.1073/pnas.0810057105
发表时间：
2008-12-16
影响因子：
11.1
作者：
Eltsov, Mikhail;MacLellan, Kirsty M.;Dubochet, Jacques
通讯作者：
Dubochet, Jacques
DOI：
10.1073/pnas.1509317112
发表时间：
2015-09-22
影响因子：
11.1
作者：
Berry, Joel;Weber, Stephanie C.;Brangwynne, Clifford P.
通讯作者：
Brangwynne, Clifford P.
影响因子：
7.8
作者：
Kireeva, N;Lakonishok, M;Belmont, AS
通讯作者：
Belmont, AS