Department of Biology, Indiana University, Bloomington, IN 47405-7003, USA.
Mol Biol Cell. 2012 Jan;23(2):284-96. doi: 10.1091/mbc.E11-09-0768. Epub 2011 Nov 30.
Cyclin B activates cyclin-dependent kinase 1 (CDK1) at mitosis, but conflicting views have emerged on the dynamics of its synthesis during embryonic cycles, ranging from continuous translation to rapid synthesis during mitosis. Here we show that a CDK1-mediated negative-feedback loop attenuates cyclin production before mitosis. Cyclin B plateaus before peak CDK1 activation, and proteasome inhibition caused minimal accumulation during mitosis. Inhibiting CDK1 permitted continual cyclin B synthesis, whereas adding nondegradable cyclin stalled it. Cycloheximide treatment before mitosis affected neither cyclin levels nor mitotic entry, corroborating this repression. Attenuated cyclin production collaborates with its destruction, since excess cyclin B1 mRNA accelerated cyclin synthesis and caused incomplete proteolysis and mitotic arrest. This repression involved neither adenylation nor the 3' untranslated region, but it corresponded with a shift in cyclin B1 mRNA from polysome to nonpolysome fractions. A pulse-driven CDK1-anaphase-promoting complex (APC) model corroborated these results, revealing reduced cyclin levels during an oscillation and permitting more effective removal. This design also increased the robustness of the oscillator, with lessened sensitivity to changes in cyclin synthesis rate. Taken together, the results of this study underscore that attenuating cyclin synthesis late in interphase improves both the efficiency and robustness of the CDK1-APC oscillator.
细胞周期蛋白 B 在有丝分裂时激活细胞周期蛋白依赖性激酶 1(CDK1),但关于其在胚胎周期中的合成动力学存在矛盾的观点,从持续翻译到有丝分裂时快速合成不等。在这里,我们表明 CDK1 介导的负反馈回路会在有丝分裂前减弱细胞周期蛋白的产生。细胞周期蛋白 B 在峰值 CDK1 激活之前达到平台期,并且蛋白酶体抑制在有丝分裂期间导致最小的积累。抑制 CDK1 允许持续的细胞周期蛋白 B 合成,而添加不可降解的细胞周期蛋白则使其停滞。有丝分裂前用环己酰亚胺处理既不影响细胞周期蛋白水平也不影响有丝分裂进入,这证实了这种抑制作用。减弱的细胞周期蛋白产生与其破坏协同作用,因为过量的细胞周期蛋白 B1 mRNA 加速了细胞周期蛋白的合成,并导致不完全的蛋白水解和有丝分裂停滞。这种抑制既不涉及腺苷酸化也不涉及 3'非翻译区,但它与细胞周期蛋白 B1 mRNA 从多核糖体转移到非多核糖体部分相对应。一个脉冲驱动的 CDK1-后期促进复合物(APC)模型证实了这些结果,显示在振荡过程中细胞周期蛋白水平降低,并允许更有效地去除。这种设计还增加了振荡器的稳健性,降低了对细胞周期蛋白合成率变化的敏感性。总之,这项研究的结果强调,在间期中晚期减弱细胞周期蛋白的合成可以提高 CDK1-APC 振荡器的效率和稳健性。