Savvidou Ellada, Cobbe Neville, Steffensen Søren, Cotterill Sue, Heck Margarete M S
Wellcome Trust Centre for Cell Biology, Institute of Cell and Molecular Biology, University of Edinburgh, Michael Swann Building, King's Buildings, Mayfield Road, Edinburgh, EH9 3JR, UK.
J Cell Sci. 2005 Jun 1;118(Pt 11):2529-43. doi: 10.1242/jcs.02392.
The precise mechanism of chromosome condensation and decondensation remains a mystery, despite progress over the last 20 years aimed at identifying components essential to the mitotic compaction of the genome. In this study, we analyse the localization and role of the CAP-D2 non-SMC condensin subunit and its effect on the stability of the condensin complex. We demonstrate that a condensin complex exists in Drosophila embryos, containing CAP-D2, the anticipated SMC2 and SMC4 proteins, the CAP-H/Barren and CAP-G (non-SMC) subunits. We show that CAP-D2 is a nuclear protein throughout interphase, increasing in level during S phase, present on chromosome axes in mitosis, and still present on chromosomes as they start to decondense late in mitosis. We analysed the consequences of CAP-D2 loss after dsRNA-mediated interference, and discovered that the protein is essential for chromosome arm and centromere resolution. The loss of CAP-D2 after RNAi has additional downstream consequences on the stability of CAP-H, the localization of DNA topoisomerase II and other condensin subunits, and chromosome segregation. Finally, we discovered that even after interfering with two components important for chromosome architecture (DNA topoisomerase II and condensin), chromosomes were still able to compact, paving the way for the identification of further components or activities required for this essential process.
尽管在过去20年里致力于确定对于基因组有丝分裂压缩至关重要的成分方面取得了进展,但染色体凝聚和去凝聚的确切机制仍是一个谜。在本研究中,我们分析了非SMC凝聚蛋白亚基CAP - D2的定位和作用及其对凝聚蛋白复合体稳定性的影响。我们证明在果蝇胚胎中存在一种凝聚蛋白复合体,其包含CAP - D2、预期的SMC2和SMC4蛋白、CAP - H/Barren和CAP - G(非SMC)亚基。我们表明CAP - D2在整个间期都是一种核蛋白,在S期水平增加,在有丝分裂时存在于染色体轴上,并且在有丝分裂后期染色体开始去凝聚时仍存在于染色体上。我们分析了dsRNA介导的干扰后CAP - D2缺失的后果,发现该蛋白对于染色体臂和着丝粒的分离至关重要。RNAi后CAP - D2的缺失对CAP - H的稳定性、DNA拓扑异构酶II和其他凝聚蛋白亚基的定位以及染色体分离有额外的下游影响。最后,我们发现即使干扰了对染色体结构重要的两个成分(DNA拓扑异构酶II和凝聚蛋白)后,染色体仍然能够压缩,这为鉴定这一关键过程所需的其他成分或活性铺平了道路。