title: Biased signaling regulates the pleiotropic effects of the urotensin II receptor to modulate its cellular behaviors.
abstract: Biased agonism by G-protein-coupled receptor ligands has opened up strategies for targeted physiological or therapeutic actions. We hypothesized that urotensin II (UII)-derived peptides displayed unexpected physiological effects because of such biased signaling on the UII human urotensin (hUT) receptor. We determined the coupling to G proteins and β-arrestins of the UII-activated hUT receptor expressed in HEK293 using bioluminescence resonance energy transfer (BRET) biosensors, as well as the production of IP1-3 and cAMP using homogenous time-resolved Forster resonance energy transfer (FRET) (HTRF)-based assays. The activated receptor coupled to Gi1, GoA, Gq, and G13, excluding Gs, and recruited β-arrestins 1 and 2. Integration of these pathways led to a 2-phase kinetic phosphorylation of ERK1/2 kinases. The tested peptides induced three different profiles: UII, urotensin-related peptide (URP), and UII4-11 displayed the full profile; [Orn(8)]UII and [Orn(5)]URP activated G proteins, although with pEC50s 5-10× higher, and did not or barely recruited β-arrestin; urantide also failed to recruit β-arrestin but displayed a reversed rank order for Gi and Gq vs. Go pEC50s (-8.79±0.20, -8.43±0.21, and -7.86±0.36, respectively, for urantide, -7.87±0.10, -7.23±0.27, and -8.55±0.19, respectively, for [Orn(5)]URP) and was a partial agonist of all G-protein pathways. Interestingly, the peptides differently modulated cell survival but similarly induced cell migration and adhesion. Thus, we demonstrate biased signaling between β-arrestin and G proteins, and between G-protein subtypes, which dictates the receptor's cellular responses.
authors.affiliation: Department of Pharmacology, Institut de Pharmacologie de Sherbrooke, Faculty of Medicine and Health Sciences, Université de Sherbrooke, Sherbrooke, Québec, Canada; Centre National de la Recherche Scientifique (CNRS), Unité Mixte de Recherche (UMR) 5203, Institut de Génomique Fonctionnelle, Montpellier, France; Institut National de la Santé et de la Recherche Médicale (INSERM), U661, Montpellier, France; UMR 5203, Universités de Montpellier 1 and 2, Montpellier, France;
authors.forename: Cédric
authors.initials: C
authors.lastname: Brulé
authors.affiliation: Department of Pharmacology, Institut de Pharmacologie de Sherbrooke, Faculty of Medicine and Health Sciences, Université de Sherbrooke, Sherbrooke, Québec, Canada; INSERM, U982, Laboratory of Neuronal and Neuroendocrine Communication and Differentiation (DC2N), Astrocyte and Vascular Niche, Biomedical Research Institute (IRIB), Pôles de Recherche et d'Enseignement Supérieur (PRES) Normandy, Peptide Research Network of Excellence (PERENE), University of Rouen, Mont-Saint-Aignan, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Perzo
authors.affiliation: INSERM, U982, Laboratory of Neuronal and Neuroendocrine Communication and Differentiation (DC2N), Astrocyte and Vascular Niche, Biomedical Research Institute (IRIB), Pôles de Recherche et d'Enseignement Supérieur (PRES) Normandy, Peptide Research Network of Excellence (PERENE), University of Rouen, Mont-Saint-Aignan, France.
authors.forename: Jane-Eileen
authors.initials: JE
authors.lastname: Joubert
authors.affiliation: Department of Pharmacology, Institut de Pharmacologie de Sherbrooke, Faculty of Medicine and Health Sciences, Université de Sherbrooke, Sherbrooke, Québec, Canada;
authors.forename: Xavier
authors.initials: X
authors.lastname: Sainsily
authors.affiliation: Department of Pharmacology, Institut de Pharmacologie de Sherbrooke, Faculty of Medicine and Health Sciences, Université de Sherbrooke, Sherbrooke, Québec, Canada;
authors.forename: Richard
authors.initials: R
authors.lastname: Leduc
authors.affiliation: INSERM, U982, Laboratory of Neuronal and Neuroendocrine Communication and Differentiation (DC2N), Astrocyte and Vascular Niche, Biomedical Research Institute (IRIB), Pôles de Recherche et d'Enseignement Supérieur (PRES) Normandy, Peptide Research Network of Excellence (PERENE), University of Rouen, Mont-Saint-Aignan, France.
authors.forename: Hélène
authors.initials: H
authors.lastname: Castel
authors.affiliation: Centre National de la Recherche Scientifique (CNRS), Unité Mixte de Recherche (UMR) 5203, Institut de Génomique Fonctionnelle, Montpellier, France; Institut National de la Santé et de la Recherche Médicale (INSERM), U661, Montpellier, France; UMR 5203, Universités de Montpellier 1 and 2, Montpellier, France; laurent.prezeau@igf.cnrs.fr.
authors.forename: Laurent
authors.initials: L
authors.lastname: Prézeau
mesh_terms: D017209:Apoptosis / Q000502:physiology; D019390:Arrestins / Q000378:metabolism; D002470:Cell Survival / Q000502:physiology; D031541:Fluorescence Resonance Energy Transfer; D019204:GTP-Binding Proteins / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D007700:Kinetics; D020935:MAP Kinase Signaling System; D010766:Phosphorylation; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism / Q000502:physiology*; D015398:Signal Transduction; D000071557:beta-Arrestins
chemical_list: D019390:Arrestins; D043562:Receptors, G-Protein-Coupled; C121697:UTS2R protein, human; D000071557:beta-Arrestins; D019204:GTP-Binding Proteins
keywords: G-protein signaling; G-protein-coupled receptor; partial agonism; β-arrestin signaling