A Clock-Binding Small Molecule Disrupts The Interaction Between Clock And Bmal1 And Enhances Circadian Rhythm Amplitude
Autor: | Yagmur Umay Doruk, Seref Gul, Ibrahim Baris, Yasemin Kubra Akyel, Narin Ozturk, Fatma Yilmaz, Alper Okyar, Darya Yarparvar, Nuri Ozturk, Metin Turkay, Ibrahim Halil Kavakli, Ali Cihan Taskin |
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Rok vydání: | 2020 |
Předmět: |
0301 basic medicine
Male Time Factors Cell Survival Circadian clock Regulator CLOCK Proteins Transcription coregulator Biology Molecular Dynamics Simulation Biochemistry Models Biological Small Molecule Libraries 03 medical and health sciences Transcription (biology) Cell Line Tumor Animals Humans Gene Regulation Circadian rhythm Molecular Biology Transcription factor Regulation of gene expression Cell Nucleus Binding Sites 030102 biochemistry & molecular biology ARNTL Transcription Factors Cell Biology Cell biology Circadian Rhythm Mice Inbred C57BL Molecular Docking Simulation Protein Transport 030104 developmental biology HEK293 Cells Liver Protein Binding Subcellular Fractions |
Zdroj: | J Biol Chem |
Popis: | Proper function of many physiological processes requires a robust circadian clock. Disruptions of the circadian clock can result in metabolic diseases, mood disorders, and accelerated aging. Therefore, identifying small molecules that specifically modulate regulatory core clock proteins may potentially enable better management of these disorders. In this study, we applied a structure-based molecular-docking approach to find small molecules that specifically bind to the core circadian regulator, the transcription factor circadian locomotor output cycles kaput (CLOCK). We identified 100 candidate molecules by virtual screening of ∼2 million small molecules for those predicted to bind closely to the interface in CLOCK that interacts with its transcriptional co-regulator, Brain and muscle Arnt-like protein-1 (BMAL1). Using a mammalian two-hybrid system, real-time monitoring of circadian rhythm in U2OS cells, and various biochemical assays, we tested these compounds experimentally and found one, named CLK8, that specifically bound to and interfered with CLOCK activity. We show that CLK8 disrupts the interaction between CLOCK and BMAL1 and interferes with nuclear translocation of CLOCK both in vivo and in vitro. Results from further experiments indicated that CLK8 enhances the amplitude of the cellular circadian rhythm by stabilizing the negative arm of the transcription/translation feedback loop without affecting period length. Our results reveal CLK8 as a tool for further studies of CLOCK's role in circadian rhythm amplitude regulation and as a potential candidate for therapeutic development to manage disorders associated with dampened circadian rhythms. |
Databáze: | OpenAIRE |
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