Marczak Marek M, Yan Bingfang
Department of Biomedical and Pharmaceutical Sciences, Center for Integrated Drug Development, University of Rhode Island, Kingston, RI 02881, United States.
Department of Biomedical and Pharmaceutical Sciences, Center for Integrated Drug Development, University of Rhode Island, Kingston, RI 02881, United States.
Arch Biochem Biophys. 2017 Oct 1;631:11-18. doi: 10.1016/j.abb.2017.08.004. Epub 2017 Aug 8.
Circadian rhythm misalignment has been increasingly recognized to pose health risk for a wide range of diseases, particularly metabolic disorders. The liver maintains metabolic homeostasis and expresses many circadian genes, such as differentiated embryo chondrocyte-1 (DEC1) and small heterodimer partner (SHP). DEC1 is established to repress transcription through E-box elements, and SHP belongs to the superfamily of nuclear receptors and has multiple E-box elements in its promoter. Importantly, DEC1 and SHP are inversely oscillated. This study was performed to test the hypothesis that the SHP gene is a target gene of DEC1. Cotransfection demonstrated that DEC1 repressed the SHP promoter and attenuated the transactivation of the classic circadian activator complex of Clock/Bmal1. Site-directed mutagenesis, electrophoretic mobility shift assay and chromatin immunoprecipitation established that the repression was achieved through the E-box in the proximal promoter. Transfection of DEC1 suppressed the expression of SHP. In circadian-inducing cells, the epileptic agent valproate inversely altered the expression of DEC1 and SHP. Both DEC1 and SHP are involved in energy balance and valproate is known to induce hepatic steatosis. Our findings collectively establish that DEC1 participates in the negative loop of SHP oscillating expression with potential implications in metabolic homeostasis.
昼夜节律失调对多种疾病,尤其是代谢紊乱所构成的健康风险已得到越来越多的认识。肝脏维持代谢稳态并表达许多昼夜节律基因,如分化胚胎软骨细胞-1(DEC1)和小异源二聚体伴侣(SHP)。DEC1被证实可通过E盒元件抑制转录,而SHP属于核受体超家族,其启动子中有多个E盒元件。重要的是,DEC1和SHP呈反向振荡。本研究旨在验证SHP基因是DEC1的靶基因这一假说。共转染表明DEC1抑制SHP启动子并减弱Clock/Bmal1经典昼夜节律激活复合物的反式激活。定点诱变、电泳迁移率变动分析和染色质免疫沉淀表明,这种抑制是通过近端启动子中的E盒实现的。DEC1转染抑制了SHP的表达。在昼夜节律诱导细胞中,癫痫药物丙戊酸反向改变了DEC1和SHP的表达。DEC1和SHP均参与能量平衡,且已知丙戊酸会诱导肝脂肪变性。我们的研究结果共同表明,DEC1参与SHP振荡表达的负反馈回路,对代谢稳态具有潜在影响。