Nuclear-cytoplasmic compartmentalization of cyclin B1-Cdk1 promotes robust timing of mitotic events
Nuclear-cytoplasmic compartmentalization of cyclin B1-Cdk1 promotes robust timing of mitotic events
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DOI：
10.1016/j.celrep.2022.111870
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发表时间：
2022-12-27
期刊：
影响因子：
8.8
通讯作者：
Yang，Qiong
中科院分区：
文献类型：
--
作者：
Maryu，Gembu;Yang，Qiong
细胞周期蛋白依赖性激酶（Cdk 1）振荡器的广泛特点是在均质胞质提取物，留下不清楚的影响，核质区室化。在这里，通过开发一个Förster共振能量转移（FRET）生物传感器，我们跟踪Cdk 1的时空动态重建细胞或没有并排，并发现区室化显着调制的时钟特性，以前发现在批量研究。虽然无核细胞显示高度可调的频率，有核细胞保持恒定的频率对细胞周期蛋白B1的变化。尽管表达变异性高，但细胞周期蛋白在相同的持续时间内降解，从而使有丝分裂期更加稳定。此外，Cdk 1和细胞周期蛋白B1严格异相循环，确保了宽的相平面轨道，这对振荡鲁棒性至关重要。虽然Cdk 1在均匀的提取物是众所周知的延迟开关样激活，我们发现活跃的细胞周期蛋白B1-Cdk 1积累在细胞核中，没有延迟，直到核膜破裂（NEB）时，另一个突然激活触发后期。cdk 1双相激活和空间区室化可能共同协调不同下游事件的准确排序。
The cyclin-dependent kinase (Cdk1) oscillator is widely characterized in homogenized cytosolic extracts, leaving unclear the impact of nucleocytoplasmic compartmentalization. Here, by developing a Förster resonance energy transfer (FRET) biosensor, we track Cdk1 spatiotemporal dynamics in reconstituted cells with or without side by side and find compartmentalization significantly modulates clock properties previously found in bulk studies. Although nucleus-absent cells display highly tunable frequency, the nucleus-present cells maintain constant frequency against cyclin B1 variations. Despite high expression variability, cyclin degraded within the same duration, enabling a robust mitotic phase. Moreover, Cdk1 and cyclin B1 cycle rigorously out-of-phase, ensuring wide phase-plane orbits, essential for oscillation robustness. Although Cdk1 in homogeneous extracts is well known for delayed switch-like activation, we find active cyclin B1-Cdk1 accumulates in nuclei, without delay, until the nuclear envelope breakdown (NEB) when another abrupt activation triggers anaphase. Cdk1 biphasic activation and spatial compartmentalization may together coordinate the accurate ordering of different downstream events.