Stable morphology, but dynamic internal reorganisation, of interphase human chromosomes in living cells.
Stable morphology, but dynamic internal reorganisation, of interphase human chromosomes in living cells.
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DOI：
10.1371/journal.pone.0011560
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发表时间：
2010-07-13
期刊：
影响因子：
3.7
通讯作者：
Chubb JR
中科院分区：
文献类型：
--
作者：
Müller I;Boyle S;Singer RH;Bickmore WA;Chubb JR
尽管有丝分裂染色体的独特结构，但还不可能在活的间期细胞中观察到单独的染色体，在那里染色体花费了90%以上的时间。间期染色体结构和动力学的研究使用固定细胞的荧光原位杂交(FISH)，这可能会破坏结构并丢失动态信息。我们开发了一种新的方法，包括在有丝分裂时对标记的组蛋白H3进行光激活，以显示活的间期细胞中的单个和特定的人类染色体。我们的数据显示，大量的染色体体积和形态在有丝分裂后迅速建立，只有在G1的第一个小时后才逐渐变化。这与标记染色体上特定基因座的行为形成对比，后者显示出更渐进的重组，并揭示了染色质从染色体区域“循环”出来是一种动态状态。我们测量了染色体解缩的相当大的异质性，甚至在姐妹染色单体之间，这可能反映了解缩的局部结构障碍，并可能放大转录噪声。染色体结构对转录、组蛋白去乙酰化和染色质重塑的抑制物表现出巨大的抵抗力。综上所述，这些数据表明，空间限制决定了结构，而不是天生的染色体结构或功能驱动的锚定，间期染色质组织主要由解缩需要和发生这种情况的可用空间之间的对立决定。
Despite the distinctive structure of mitotic chromosomes, it has not been possible to visualise individual chromosomes in living interphase cells, where chromosomes spend over 90% of their time. Studies of interphase chromosome structure and dynamics use fluorescence in-situ hybridisation (FISH) on fixed cells, which potentially damages structure and loses dynamic information. We have developed a new methodology, involving photoactivation of labelled histone H3 at mitosis, to visualise individual and specific human chromosomes in living interphase cells. Our data revealed bulk chromosome volume and morphology are established rapidly after mitosis, changing only incrementally after the first hour of G1. This contrasted with the behaviour of specific loci on labelled chromosomes, which showed more progressive reorganisation, and revealed that “looping out” of chromatin from chromosome territories is a dynamic state. We measured considerable heterogeneity in chromosome decondensation, even between sister chromatids, which may reflect local structural impediments to decondensation and could potentially amplify transcriptional noise. Chromosome structure showed tremendous resistance to inhibitors of transcription, histone deacetylation and chromatin remodelling. Together, these data indicate steric constraints determine structure, rather than innate chromosome architecture or function-driven anchoring, with interphase chromatin organisation governed primarily by opposition between needs for decondensation and the space available for this to happen.
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在鱼类实验中保存大规模染色质结构。
影响因子：
1.6
作者：
Hepperger, Claudia;Otten, Simone;von Hase, Johann;Dietzel, Steffen
通讯作者：
Dietzel, Steffen
DOI：
10.1083/jcb.145.6.1119
发表时间：
1999-06-14
期刊：
The Journal of cell biology
影响因子：
--
作者：
Croft JA;Bridger JM;Boyle S;Perry P;Teague P;Bickmore WA
通讯作者：
Bickmore WA
影响因子：
5.3
作者：
Goetze, Sandra;Mateos-Langerak, Julio;van Driel, Roel
通讯作者：
van Driel, Roel
影响因子：
9.8
作者：
Branco MR;Pombo A
通讯作者：
Pombo A
影响因子：
9.8
作者：
Ferrai C;Xie SQ;Luraghi P;Munari D;Ramirez F;Branco MR;Pombo A;Crippa MP
通讯作者：
Crippa MP