Cyclin-Dependent Kinase Sensor Transgenic Zebrafish Lines for Improved Cell Cycle State Visualization in Live Animals.
Cyclin-Dependent Kinase Sensor Transgenic Zebrafish Lines for Improved Cell Cycle State Visualization in Live Animals.
复制标题
细胞周期蛋白依赖性激酶传感器转基因斑马鱼系可改善活体动物细胞周期状态的可视化。
DOI：
10.1089/zeb.2021.0059
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发表时间：
2021
期刊：
影响因子：
2
通讯作者：
Martin,BenjaminL
中科院分区：
文献类型：
--
作者：
Morabito,RobertD;Adikes,RebeccaC;Matus,DavidQ;Martin,BenjaminL
Adikes等人。1描述了一种新型的斑马鱼细胞周期传感器，该传感器基于细胞周期蛋白依赖性激酶(CDK)的活性水平来描绘细胞周期的所有阶段。CDK传感器由人DNA解旋酶B(DHB)片段与荧光蛋白融合而成。2DHB含有一个显性核定位序列(NLS)和一个核输出序列(NES)，其两侧是CDK特异性的磷酸化位点。含有低水平CDK活性的细胞有一个暴露的NLS，它将CDK传感器定位到细胞核。随着细胞周期的进展，CDK活性增加，导致DHB的磷酸化，从而阻断NLS，并允许NES促进传感器输出到细胞质(图1)。作者发现，该比率传感器的定量分析可以区分CDK低(CDKlow)、静止的G0期细胞和CDK增加(CDKINC)、周期G1期的细胞，以及识别细胞周期的S、G2和M期。
Adikes et al. 1 described a novel zebrafish cell cycle sensor that delineates all phases of the cell cycle based on levels of cyclin-dependent kinase (CDK) activity. The CDK sensor consists of a fragment of human DNA Helicase B (DHB) fused to a fluorescent protein. 2 DHB contains a dominant nuclear localization sequence (NLS) and a nuclear export sequence (NES) flanked by CDK-specific phosphorylation sites. Cells that contain low levels of CDK activity have an exposed NLS that localizes the CDK sensor to the nucleus. As the cell cycle progresses, CDK activity increases causing phosphorylation of DHB, which occludes the NLS and allows the NES to promote export of the sensor into the cytoplasm (Fig. 1). The authors showed that quantitative analysis of this ratiometric sensor can distinguish CDK low (CDKlow), quiescent G0 arrested cells from CDK increasing (CDKinc), cycling G1 phase cells, as well as identify S, G2, and M phases of the cell cycle.