Casein kinase 1 proteomics reveal prohibitin 2 function in molecular clock.
Casein kinase 1 proteomics reveal prohibitin 2 function in molecular clock.
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DOI：
10.1371/journal.pone.0031987
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Fu YH
中科院分区：
文献类型：
--
作者：
Kategaya LS;Hilliard A;Zhang L;Asara JM;Ptáček LJ;Fu YH
在一天中，时钟蛋白同步动物生理学的变化（例如，觉醒和食欲）与外部线索（例如，日光和食物）。在脊椎动物中，酪蛋白激酶1 δ和β（CK1δ和CK1ε）通过磷酸化其他核心时钟蛋白来调节这些昼夜变化。此外，CK1还能以非催化方式调节昼夜节律依赖的转录，但其机制尚不清楚。此外，这些密切相关的激酶之间的功能冗余的程度是有争议的。为了进一步了解CK1δ和CK1ε在生物钟中的作用机制，我们首先对人类细胞中的两种激酶进行了蛋白质组学分析。接下来，我们在基于细胞的昼夜节律读出中测试了感兴趣的候选者，这导致了抑制蛋白2（PHB2）作为周期长度调节剂的发现。降低PHB2的表达增加了昼夜节律驱动的转录，从而揭示了PHB2在分子钟中起抑制剂的作用。虽然没有检测到PHB2与任一激酶的稳定结合，但敲低CK1ε表达增加PHB2蛋白水平，并且出乎意料地，敲低CK1δ降低PHB2转录物水平。因此，分离CK1蛋白复合物导致鉴定PHB2作为昼夜节律转录的抑制剂。此外，我们发现CK1δ和CK1ε差异调节PHB2的表达。
Throughout the day, clock proteins synchronize changes in animal physiology (e.g., wakefulness and appetite) with external cues (e.g., daylight and food). In vertebrates, both casein kinase 1 delta and epsilon (CK1δ and CK1ε) regulate these circadian changes by phosphorylating other core clock proteins. In addition, CK1 can regulate circadian-dependent transcription in a non-catalytic manner, however, the mechanism is unknown. Furthermore, the extent of functional redundancy between these closely related kinases is debated. To further advance knowledge about CK1δ and CK1ε mechanisms of action in the biological clock, we first carried out proteomic analysis of both kinases in human cells. Next, we tested interesting candidates in a cell-based circadian readout which resulted in the discovery of PROHIBITIN 2 (PHB2) as a modulator of period length. Decreasing the expression of PHB2 increases circadian-driven transcription, thus revealing PHB2 acts as an inhibitor in the molecular clock. While stable binding of PHB2 to either kinase was not detected, knocking down CK1ε expression increases PHB2 protein levels and, unexpectedly, knocking down CK1δ decreases PHB2 transcript levels. Thus, isolating CK1 protein complexes led to the identification of PHB2 as an inhibitor of circadian transcription. Furthermore, we show that CK1δ and CK1ε differentially regulate the expression of PHB2.
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影响因子：
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