Doughty Tyler W, Arsenault Heather E, Benanti Jennifer A
Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, Worcester, Massachusetts, United States of America.
PLoS Genet. 2016 Jul 27;12(7):e1006216. doi: 10.1371/journal.pgen.1006216. eCollection 2016 Jul.
During mitosis chromosomes are condensed to facilitate their segregation, through a process mediated by the condensin complex. Although several factors that promote maximal condensin activity during mitosis have been identified, the mechanisms that downregulate condensin activity during interphase are largely unknown. Here, we demonstrate that Ycg1, the Cap-G subunit of budding yeast condensin, is cell cycle-regulated with levels peaking in mitosis and decreasing as cells enter G1 phase. This cyclical expression pattern is established by a combination of cell cycle-regulated transcription and constitutive degradation. Interestingly, overexpression of YCG1 and mutations that stabilize Ycg1 each result in delayed cell-cycle entry and an overall proliferation defect. Overexpression of no other condensin subunit impacts the cell cycle, suggesting that Ycg1 is limiting for condensin complex formation. Consistent with this possibility, we find that levels of intact condensin complex are reduced in G1 phase compared to mitosis, and that increased Ycg1 expression leads to increases in both levels of condensin complex and binding to chromatin in G1. Together, these results demonstrate that Ycg1 levels limit condensin function in interphase cells, and suggest that the association of condensin with chromosomes must be reduced following mitosis to enable efficient progression through the cell cycle.
在有丝分裂期间,染色体通过凝缩素复合体介导的过程进行浓缩,以促进其分离。尽管已经鉴定出几种在有丝分裂期间促进凝缩素活性达到最大值的因子,但在间期下调凝缩素活性的机制在很大程度上仍不清楚。在这里,我们证明芽殖酵母凝缩素的Cap-G亚基Ycg1受细胞周期调控,其水平在有丝分裂时达到峰值,并随着细胞进入G1期而下降。这种周期性表达模式是由细胞周期调控转录和组成型降解共同建立的。有趣的是,YCG1的过表达以及使Ycg1稳定的突变均导致细胞周期进入延迟和整体增殖缺陷。其他凝缩素亚基的过表达均不影响细胞周期,这表明Ycg1对凝缩素复合体的形成具有限制作用。与此可能性一致,我们发现与有丝分裂相比,G1期完整凝缩素复合体的水平降低,并且Ycg1表达增加导致G1期凝缩素复合体水平及其与染色质结合的增加。总之,这些结果表明Ycg1水平限制了间期细胞中凝缩素的功能,并表明有丝分裂后凝缩素与染色体的结合必须减少,以实现细胞周期的有效进程。