Optogenetic interrogation reveals separable G-protein-dependent and -independent signalling linking G-protein-coupled receptors to the circadian oscillator

BMC Biology

Research Article

Takeaki Ozawa¹  Tomoki Nishiguchi¹  Helena J. Bailes²  Nina Milosavljevic²  Elliot J. Gerrard²  Robert J. Lucas²  Ling-Yu Zhuang² 

[1] Department of Chemistry, The University of Tokyo, Tokyo, Japan;Faculty of Biology, Medicine and Health, The University of Manchester, Manchester, UK;

关键词: HEK293 Cell;    Circadian Clock;    Phase Response Curve;    Proximity Ligation Assay;    Rat1 Fibroblast;   

DOI  :  10.1186/s12915-017-0380-8

 received in 2017-01-18, accepted in 2017-04-26,  发布年份 2017

来源: Springer

【 摘 要 】

BackgroundEndogenous circadian oscillators distributed across the mammalian body are synchronised among themselves and with external time via a variety of signalling molecules, some of which interact with G-protein-coupled receptors (GPCRs). GPCRs can regulate cell physiology via pathways originating with heterotrimeric G-proteins or β-arrestins. We applied an optogenetic approach to determine the contribution of these two signalling modes on circadian phase.ResultsWe employed a photopigment (JellyOp) that activates Gαs signalling with better selectivity and higher sensitivity than available alternatives, and a point mutant of this pigment (F112A) biased towards β-arrestin signalling. When expressed in fibroblasts, both native JellyOp and the F112A arrestin-biased mutant drove light-dependent phase resetting in the circadian clock. Shifts induced by the two opsins differed in their circadian phase dependence and the degree to which they were associated with clock gene induction.ConclusionsOur data imply separable G-protein and arrestin inputs to the mammalian circadian clock and establish a pair of optogenetic tools suitable for manipulating Gαs- and β-arrestin-biased signalling in live cells.

【 授权许可】

CC BY   
© Milosavljevic et al. 2017