Expression Levels of Estrogen Receptor β Are Modulated by Components of the Molecular Clock

Abstract

Circadian regulation of gene expression plays a major role in health and disease. The precise role of the circadian system remains to be clarified, but it is known that circadian proteins generate physiological rhythms in organisms by regulating clock-controlled target genes. The estrogen receptor beta (ERβ) is, together with ERα, a member of the nuclear receptor superfamily and a key mediator of estrogen action. Interestingly, recent studies show that disturbed circadian rhythmicity in humans can increase the risk of reproductive malfunctions, suggesting a link between the circadian system and ER-mediated transcription pathways. Here, we identify a novel level of regulation of estrogen signaling where ERβ, but not ERα, is controlled by circadian clock proteins. We show that ERβ mRNA levels fluctuate in different peripheral tissues following a robust circadian pattern, with a peak at the light-dark transition, which is maintained under free-running conditions. Interestingly, this oscillation is abolished in clock-deficient BMAL1 knockout mice. Circadian control of ERβ expression is exerted through a conserved E-box element in the ERβ promoter region that recruits circadian regulatory factors. Furthermore, using small interfering RNA-mediated knockdown assays, we show that the expression levels of the circadian regulatory factors directly influence estrogen signaling by regulating the intracellular levels of endogenous ERβ.

ACKNOWLEDGMENTS

This work was supported by the European Union thorough the CASCADE Network of Excellence and the CRESCENDO Integrated Project, by the Swedish Cancer Fund, by the Swedish Research Council, by the CNRS, the MENRT, the Association pour la Recherche sur le Cancer, and by Université de Nice-Sophia-Antipolis.

We thank Gerard Triqueneaux for technical help.

Jan-Åke Gustafsson is a shareholder, research grant recipient, and consultant of KaroBio AB.