Biased signaling through G-protein-coupled PROKR2 receptors harboring missense mutations.
Various missense mutations in the gene coding for prokineticin receptor 2 (PROKR2), a G-protein-coupled receptor, have been identified in patients with Kallmann syndrome. However, the functional consequences of these mutations on the different signaling pathways of this receptor have not been studied. We first showed that the wild-type PROKR2 can activate different G-protein subtypes (Gq, Gs, and Gi/o) and recruit β-arrestins in transfected HEK-293 cells. We then examined, for each of these signaling pathways, the effects of 9 mutations that did not significantly impair cell surface targeting or ligand binding of the receptor. Four mutant receptors showing defective Gq signaling (R85C, R85H, R164Q, and V331M) could still recruit β-arrestins on ligand activation, which may cause biased signaling in vivo. Conversely, the R80C receptor could activate the 3 types of G proteins but could not recruit β-arrestins. Finally, the R268C receptor could recruit β-arrestins and activate the Gq and Gs signaling pathways but could not activate the Gi/o signaling pathway. Our results validate the concept that mutations in the genes encoding membrane receptors can bias downstream signaling in various ways, possibly leading to pathogenic and, perhaps in some cases, protective (e.g., R268C) effects.
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; Université Montpellier 1 and 2, Montpellier, France;
Oualid
O
Sbai
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; Université Montpellier 1 and 2, Montpellier, France;
Carine
C
Monnier
EA7331, Faculté des Sciences Pharmaceutiques et Biologiques, Université Paris-Descartes, Paris, France; and.
Catherine
C
Dodé
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; Université Montpellier 1 and 2, Montpellier, France;
Jean-Philippe
JP
Pin
INSERM Unité Mixte de Recherche en Santé (UMRS) 1120, Département Neuroscience, Institut Pasteur, Paris, France philippe.rondard@igf.cnrs.fr jean-pierre.hardelin@pasteur.fr.
Jean-Pierre
JP
Hardelin
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; Université Montpellier 1 and 2, Montpellier, France; philippe.rondard@igf.cnrs.fr jean-pierre.hardelin@pasteur.fr.
Philippe
P
Rondard
D019390:Arrestins / Q000378:metabolism*; D020013:Calcium Signaling / Q000235:genetics / Q000502:physiology; D019206:GTP-Binding Protein alpha Subunits, Gi-Go / Q000378:metabolism*; D043802:GTP-Binding Protein alpha Subunits, Gq-G11 / Q000378:metabolism*; D019205:GTP-Binding Protein alpha Subunits, Gs / Q000378:metabolism*; D005768:Gastrointestinal Hormones / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D017436:Kallmann Syndrome / Q000235:genetics*; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D020125:Mutation, Missense; D009479:Neuropeptides / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D021381:Protein Transport; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000502:physiology*; D018000:Receptors, Peptide / Q000235:genetics / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015398:Signal Transduction / Q000235:genetics / Q000502:physiology*; D014162:Transfection; D000071557:beta-Arrestins
D019390:Arrestins; D005768:Gastrointestinal Hormones; D009479:Neuropeptides; C423608:PROK2 protein, human; C470838:PROKR2 protein, human; D043562:Receptors, G-Protein-Coupled; D018000:Receptors, Peptide; D011993:Recombinant Fusion Proteins; D000071557:beta-Arrestins; D019206:GTP-Binding Protein alpha Subunits, Gi-Go; D043802:GTP-Binding Protein alpha Subunits, Gq-G11; D019205:GTP-Binding Protein alpha Subunits, Gs
Kallmann syndrome; prokineticin; β-arrestin