title: USP2-45 Is a Circadian Clock Output Effector Regulating Calcium Absorption at the Post-Translational Level.
abstract: The mammalian circadian clock influences most aspects of physiology and behavior through the transcriptional control of a wide variety of genes, mostly in a tissue-specific manner. About 20 clock-controlled genes (CCGs) oscillate in virtually all mammalian tissues and are generally considered as core clock components. One of them is Ubiquitin-Specific Protease 2 (Usp2), whose status remains controversial, as it may be a cogwheel regulating the stability or activity of core cogwheels or an output effector. We report here that Usp2 is a clock output effector related to bodily Ca2+ homeostasis, a feature that is conserved across evolution. Drosophila with a whole-body knockdown of the orthologue of Usp2, CG14619 (dUsp2-kd), predominantly die during pupation but are rescued by dietary Ca2+ supplementation. Usp2-KO mice show hyperabsorption of dietary Ca2+ in small intestine, likely due to strong overexpression of the membrane scaffold protein NHERF4, a regulator of the Ca2+ channel TRPV6 mediating dietary Ca2+ uptake. In this tissue, USP2-45 is found in membrane fractions and negatively regulates NHERF4 protein abundance in a rhythmic manner at the protein level. In clock mutant animals (Cry1/Cry2-dKO), rhythmic USP2-45 expression is lost, as well as the one of NHERF4, confirming the inverse relationship between USP2-45 and NHERF4 protein levels. Finally, USP2-45 interacts in vitro with NHERF4 and endogenous Clathrin Heavy Chain. Taken together these data prompt us to define USP2-45 as the first clock output effector acting at the post-translational level at cell membranes and possibly regulating membrane permeability of Ca2+.
authors.affiliation: University of Lausanne, Department of Pharmacology and Toxicology, Rue du Bugnon 27, CH-1005 Lausanne, Switzerland.
authors.forename: Daniel
authors.initials: D
authors.lastname: Pouly
authors.affiliation: University of Lausanne, Department of Pharmacology and Toxicology, Rue du Bugnon 27, CH-1005 Lausanne, Switzerland.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Chenaux
authors.affiliation: University of Lausanne, Department of Pharmacology and Toxicology, Rue du Bugnon 27, CH-1005 Lausanne, Switzerland.
authors.forename: Virginie
authors.initials: V
authors.lastname: Martin
authors.affiliation: University of Lausanne, Department of Pharmacology and Toxicology, Rue du Bugnon 27, CH-1005 Lausanne, Switzerland.
authors.forename: Maja
authors.initials: M
authors.lastname: Babis
authors.affiliation: University of Geneva, Department of Genetics & Evolution, 30, Quai Ernest Ansermet, CH-1211 Geneva, Switzerland.
authors.forename: Rafael
authors.initials: R
authors.lastname: Koch
authors.affiliation: University of Geneva, Department of Genetics & Evolution, 30, Quai Ernest Ansermet, CH-1211 Geneva, Switzerland.
authors.forename: Emi
authors.initials: E
authors.lastname: Nagoshi
authors.affiliation: University of Lausanne, Department of Pharmacology and Toxicology, Rue du Bugnon 27, CH-1005 Lausanne, Switzerland.
authors.forename: Vladimir L
authors.initials: VL
authors.lastname: Katanaev
authors.affiliation: University of Lausanne, Department of Pharmacology and Toxicology, Rue du Bugnon 27, CH-1005 Lausanne, Switzerland.|Department of Diabetes and Circadian Rhythms, Nestlé Institute of Health Sciences, CH-1015 Lausanne, Switzerland.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Gachon
authors.affiliation: University of Lausanne, Department of Pharmacology and Toxicology, Rue du Bugnon 27, CH-1005 Lausanne, Switzerland.|National Centre of Competence in Research "Kidney.ch", Zurich, Switzerland.
authors.forename: Olivier
authors.initials: O
authors.lastname: Staub
mesh_terms: D065567:Absorption, Physiological; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D057906:Circadian Clocks; D033922:Clathrin Heavy Chains / Q000378:metabolism; D056931:Cryptochromes / Q000378:metabolism; D004331:Drosophila melanogaster / Q000378:metabolism; D057809:HEK293 Cells; D006706:Homeostasis; D006801:Humans; D053565:Hypercalciuria / Q000378:metabolism; D007413:Intestinal Mucosa / Q000378:metabolism; D008124:Locomotion; D008297:Male; D008566:Membranes / Q000378:metabolism; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008954:Models, Biological; D010750:Phosphoproteins / Q000378:metabolism; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D017923:Sodium-Hydrogen Exchangers / Q000378:metabolism; D043222:Ubiquitin Thiolesterase; D064570:Ubiquitin-Specific Proteases / Q000378:metabolism*; D015854:Up-Regulation
chemical_list: D056931:Cryptochromes; D010750:Phosphoproteins; D017923:Sodium-Hydrogen Exchangers; C093376:sodium-hydrogen exchanger regulatory factor; D033922:Clathrin Heavy Chains; D043222:Ubiquitin Thiolesterase; D064570:Ubiquitin-Specific Proteases; C579224:Usp2 protein, mouse; D002118:Calcium