3,4-Dibromo-7-Azaindole Modulates Arabidopsis Circadian Clock by Inhibiting Casein Kinase 1 Activity
3,4-Dibromo-7-Azaindole Modulates Arabidopsis Circadian Clock by Inhibiting Casein Kinase 1 Activity
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DOI：
10.1093/pcp/pcz183
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发表时间：
2019-11-01
影响因子：
4.9
通讯作者：
Nakamichi, Norihito
中科院分区：
文献类型：
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作者：
Ono, Azusa;Sato, Ayato;Nakamichi, Norihito
生物钟是一个计时系统，用于调节许多生物的日常节奏。昼夜节律钟的一个特征是，尽管环境波动，如温度变化，周期长度仍保持相对恒定。本研究利用ITbM化学文库（ITbM chemical library）收集的小分子资料，发现3,4-二溴-7-叠氮素（B-AZ）可以延长拟南芥（Arabidopsis thaliana）的生理周期。B-AZ以前未被报道有任何生物和生化活性。靶标鉴定可以阐明小分子的作用方式，但我们无法制作B-AZ分子探针进行靶标鉴定。相反，我们进行了其他分析，基因表达谱，潜在地揭示了分子的作用模式。短期治疗B-AZ降低了4个黎明和早晨生物钟相关基因的表达，即生物钟相关基因1 （CCA1）、晚拉长下胚轴（LHY）、伪反应调节因子9 （PRR9）和PRR7。同样，B-AZ处理后，PRR5的量和CAB表达1 （TOC1）蛋白、CCA1、LHY、PRR9和PRR7的转录抑制因子的量和时间都增加了。B-AZ抑制酪蛋白激酶1家族（CK1）磷酸化PRR5和TOC1进行靶向降解。对接研究和分子动力学模拟表明，B-AZ与人类CK1三角洲的atp结合口袋相互作用，其氨基酸序列与拟南芥CK1高度相似。b - az诱导的周期延长效应在prr5 - c1突变体中减弱。总之，本研究提供了一种结构简单的新型CK1抑制剂，通过PRR5和TOC1的积累来调节生物钟。
The circadian clock is a timekeeping system for regulation of numerous biological daily rhythms. One characteristic of the circadian clock is that period length remains relatively constant in spite of environmental fluctuations, such as temperature change. Here, using the curated collection of in-house small molecule chemical library (ITbM chemical library), we show that small molecule 3,4-dibromo-7-azaindole (B-AZ) lengthened the circadian period of Arabidopsis thaliana (Arabidopsis). B-AZ has not previously been reported to have any biological and biochemical activities. Target identification can elucidate the mode of action of small molecules, but we were unable to make a molecular probe of B-AZ for target identification. Instead, we performed other analysis, gene expression profiling that potentially reveals mode of action of molecules. Short-term treatment of B-AZ decreased the expression of four dawn- and morning-phased clock-associated genes, CIRCADIAN CLOCK-ASSOCIATED 1 (CCA1), LATE ELONGATED HYPOCOTYL (LHY), PSEUDO-RESPONSE REGULATOR 9 (PRR9) and PRR7. Consistently, amounts of PRR5 and TIMING OF CAB EXPRESSION 1 (TOC1) proteins, transcriptional repressors of CCA1, LHY, PRR9 and PRR7 were increased upon B-AZ treatment. B-AZ inhibited Casein Kinase 1 family (CK1) that phosphorylates PRR5 and TOC1 for targeted degradation. A docking study and molecular dynamics simulation suggested that B-AZ interacts with the ATP-binding pocket of human CK1 delta, whose amino acid sequences are highly similar to those of Arabidopsis CK1. B-AZ-induced period-lengthening effect was attenuated in prr5 toc1 mutants. Collectively, this study provides a novel and simple structure CK1 inhibitor that modulates circadian clock via accumulation of PRR5 and TOC1.