Functional analysis after rapid degradation of condensins and 3D-EM reveals chromatin volume is uncoupled from chromosome architecture in mitosis.
Functional analysis after rapid degradation of condensins and 3D-EM reveals chromatin volume is uncoupled from chromosome architecture in mitosis.
复制标题
凝缩蛋白快速降解和 3D-EM 后的功能分析表明，染色质体积与有丝分裂中的染色体结构脱钩。
DOI：
10.1242/jcs.210187
复制
发表时间：
2018-02-22
影响因子：
4
通讯作者：
Earnshaw WC
中科院分区：
文献类型：
--
作者：
Samejima K;Booth DG;Ogawa H;Paulson JR;Xie L;Watson CA;Platani M;Kanemaki MT;Earnshaw WC
体细胞染色体形成对凝缩蛋白的需求仍不清楚，因为通过常规方法耗尽凝缩蛋白的细胞中确实会形成不完全凝缩的染色体。为了剖析凝缩素在脊椎动物有丝分裂不同阶段的作用，我们建立了一个多功能细胞系统，将生长素介导的快速降解与化学遗传学相结合，以获得鸡DT40细胞在存在和不存在凝缩素的情况下近乎同步的有丝分裂进入。我们通过活细胞和固定细胞显微镜方法分析了结果，包括具有数字重建的串行块面扫描电子显微镜。凝缩蛋白快速耗尽后，染色体缺陷比凝缩蛋白缓慢耗尽后明显得多。有丝分裂染色质总体积与对照细胞相似，但有丝分裂染色体的单个质量聚集在弯曲的有丝分裂纺锤体的一侧。培养物在前中期停滞，最终退出有丝分裂而不分离染色体。滴定生长素浓度的实验表明，后期染色体分离和正常染色体结构的形成需要不同的凝缩素水平。本文有对该论文第一作者的相关第一人称采访。摘要：凝缩蛋白的快速消耗揭示了有丝分裂染色体结构和分离需要不同的凝缩蛋白水平。有丝分裂期间染色质体积压缩不需要凝缩蛋白。
The requirement for condensin in chromosome formation in somatic cells remains unclear, as imperfectly condensed chromosomes do form in cells depleted of condensin by conventional methodologies. In order to dissect the roles of condensin at different stages of vertebrate mitosis, we have established a versatile cellular system that combines auxin-mediated rapid degradation with chemical genetics to obtain near-synchronous mitotic entry of chicken DT40 cells in the presence and absence of condensin. We analyzed the outcome by live- and fixed-cell microscopy methods, including serial block face scanning electron microscopy with digital reconstruction. Following rapid depletion of condensin, chromosomal defects were much more obvious than those seen after a slow depletion of condensin. The total mitotic chromatin volume was similar to that in control cells, but a single mass of mitotic chromosomes was clustered at one side of a bent mitotic spindle. Cultures arrest at prometaphase, eventually exiting mitosis without segregating chromosomes. Experiments where the auxin concentration was titrated showed that different condensin levels are required for anaphase chromosome segregation and formation of a normal chromosome architecture. This article has an associated First Person interview with the first author of the paper. Summary: Rapid condensin depletion reveals that different condensin levels are required for mitotic chromosome architecture and segregation. Condensin is not required for chromatin volume compaction during mitosis.
登录
查看更多内容
影响因子：
21.3
作者：
Houlard, Martin;Godwin, Jonathan;Metson, Jean;Lee, Jibak;Hirano, Tatsuya;Nasmyth, Kim
通讯作者：
Nasmyth, Kim
影响因子：
16
作者：
Booth, Daniel G.;Beckett, Alison J.;Molina, Oscar;Samejima, Itaru;Masumoto, Hiroshi;Kouprina, Natalay;Larionov, Vladimir;Prior, Ian A.;Earnshaw, William C.
通讯作者：
Earnshaw, William C.
影响因子：
15
作者：
Bishop, AC;Kung, CY;Liu, Y
通讯作者：
Liu, Y
影响因子：
64.5
作者：
GASSER, SM;LAEMMLI, UK
通讯作者：
LAEMMLI, UK
DOI：
10.1039/p19960001545
发表时间：
1996-07-07
期刊：
JOURNAL OF THE CHEMICAL SOCIETY-PERKIN TRANSACTIONS 1
影响因子：
--
作者：
Hanefeld, U;Rees, CW;Williams, DJ
通讯作者：
Williams, DJ