A chemical biology approach reveals period shortening of the mammalian circadian clock by specific inhibition of GSK-3β
A chemical biology approach reveals period shortening of the mammalian circadian clock by specific inhibition of GSK-3β
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DOI：
10.1073/pnas.0811410106
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发表时间：
2008-12-30
影响因子：
11.1
通讯作者：
Kay, Steve A.
中科院分区：
文献类型：
--
作者：
Hirota, Tsuyoshi;Lewis, Warren G.;Kay, Steve A.
生物钟在细胞水平上控制基因表达的每日振荡。我们报告了一个高通量的昼夜节律功能测定系统，包括发光报告细胞，筛选自动化和数据分析管道的发展。我们应用这个系统，进一步剖析哺乳动物生物钟的分子机制，使用化学生物学的方法。我们分析了1，280种具有不同结构的生物活性化合物对昼夜节律周期长度的影响，这表明了核心时钟机制。我们的筛选模式鉴定了许多以前已知的改变昼夜节律周期或相位的化合物，证明了测定系统的有效性。此外，我们发现，糖原合成酶激酶3（GSK-3）的小分子抑制剂一贯引起强烈的短周期表型相反，众所周知的延长期锂，另一个假定的GSK-3抑制剂。GSK-3 β的siRNA介导的敲低也引起短时间，证实了用小分子抑制剂获得的表型。这些结果阐明了GSK-3 β在哺乳动物生物钟周期调节中的作用，并强调了化学生物学在探索生物钟未知机制方面的有效性。
The circadian clock controls daily oscillations of gene expression at the cellular level. We report the development of a high-throughput circadian functional assay system that consists of luminescent reporter cells, screening automation, and a data analysis pipeline. We applied this system to further dissect the molecular mechanisms underlying the mammalian circadian clock using a chemical biology approach. We analyzed the effect of 1,280 pharmacologically active compounds with diverse structures on the circadian period length that is indicative of the core clock mechanism. Our screening paradigm identified many compounds previously known to change the circadian period or phase, demonstrating the validity of the assay system. Furthermore, we found that small molecule inhibitors of glycogen synthase kinase 3 (GSK-3) consistently caused a strong short period phenotype in contrast to the well-known period lengthening by lithium, another presumed GSK-3 inhibitor. siRNA-mediated knockdown of GSK-3 beta also caused a short period, confirming the phenotype obtained with the small molecule inhibitors. These results clarify the role of GSK-3 beta in the period regulation of the mammalian clockworks and highlight the effectiveness of chemical biology in exploring unidentified mechanisms of the circadian clock.