Gerlich Daniel, Hirota Toru, Koch Birgit, Peters Jan-Michael, Ellenberg Jan
Gene Expression and Cell Biology/Biophysics Programmes, EMBL, 69117 Heidelberg, Germany.
Curr Biol. 2006 Feb 21;16(4):333-44. doi: 10.1016/j.cub.2005.12.040.
Restructuring chromatin into morphologically distinct chromosomes is essential for cell division, but the molecular mechanisms underlying this process are poorly understood. Condensin complexes have been proposed as key factors, although controversial conclusions about their contribution to chromosome structure were reached by different experimental approaches in fixed cells or cell extracts. Their function under physiological conditions still needs to be defined.
Here, we investigated the specific functions of condensin I and II in live cells by fluorescence microscopy and RNAi depletion. Photobleaching and quantitative time-lapse imaging showed that GFP-tagged condensin II bound stably to chromosomes throughout mitosis. By contrast, the canonical condensin I interacted dynamically with chromatin after completion of prophase compaction, reaching steady-state levels on chromosomes before congression. In condensin I-depleted cells, compaction was normal, but chromosomes were mechanically labile and unable to withstand spindle forces during alignment. However, normal levels of condensin II were not required for chromosome stability.
We conclude that while condensin I seems dispensable for normal chromosome compaction, its dynamic binding after nuclear envelope breakdown locks already condensed chromatin in a rigid state required for mechanically stable spindle attachment.
将染色质重构成形态上不同的染色体对细胞分裂至关重要,但这一过程背后的分子机制仍知之甚少。凝聚素复合体被认为是关键因素,尽管在固定细胞或细胞提取物中通过不同的实验方法得出了关于它们对染色体结构贡献的有争议的结论。它们在生理条件下的功能仍有待确定。
在这里,我们通过荧光显微镜和RNAi缺失研究了活细胞中凝聚素I和II的特定功能。光漂白和定量延时成像显示,绿色荧光蛋白标记的凝聚素II在整个有丝分裂过程中稳定地与染色体结合。相比之下,典型的凝聚素I在前期压缩完成后与染色质动态相互作用,在染色体排列之前达到稳态水平。在凝聚素I缺失的细胞中,压缩正常,但染色体在机械上不稳定,在排列过程中无法承受纺锤体的力量。然而,染色体稳定性并不需要正常水平的凝聚素II。
我们得出结论,虽然凝聚素I似乎对于正常的染色体压缩是可有可无的,但其在核膜破裂后的动态结合将已经凝聚的染色质锁定在机械稳定的纺锤体附着所需的刚性状态。