This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Copyright Permission Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Ferrari, S. L. Articles by Bisello, A. Search for Related Content PubMed PubMed Citation Articles by Ferrari, S. L. Articles by Bisello, A.

Cellular Distribution of Constitutively Active Mutant Parathyroid Hormone (PTH)/PTH-Related Protein Receptors and Regulation of Cyclic Adenosine 3',5'-Monophosphate Signaling by ß-Arrestin2

Division of Bone and Mineral Metabolism Harvard-Thorndike and Charles A. Dana Research Laboratories Department of Medicine Beth Israel Deaconess Medical Center and Harvard Medical School Boston, Massachusetts 02215

PTH promotes endocytosis of human PTH receptor 1 (PTH1Rc) by activating protein kinase C and recruiting ß-arrestin2. We examined the role of ß-arrestin2 in regulating the cellular distribution and cAMP signaling of two constitutively active PTH1Rc mutants, H223R and T410P. Overexpression of a ß-arrestin2-green fluorescent protein (GFP) conjugate in COS-7 cells inhibited constitutive cAMP accumulation by H223R and T410P in a dose-dependent manner, as well as the response to PTH of both mutant and wild-type PTH1Rcs. The cellular distribution of PTH1Rc-GFP conjugates, fluorescent ligands, and ßarrestin2-GFP was analyzed by fluorescence microscopy in HEK-293T cells. In cells expressing either receptor mutant, a ligand-independent mobilization of ß-arrestin2 to the cell membrane was observed. In the absence of ligand, H223R and wild-type PTH1Rcs were mainly localized on the cell membrane, whereas intracellular trafficking of T410P was also observed. While agonists promoted ß-arrestin2-mediated endocytosis of both PTH1Rc mutants, antagonists were rapidly internalized only with T410P. The protein kinases inhibitor, staurosporine, significantly decreased internalization of ligand-PTH1Rc mutant complexes, although the recruitment of ß-arrestin2 to the cell membrane was unaffected. Moreover, in cells expressing a truncated wild-type PTH1Rc lacking the C-terminal cytoplasmic domain, agonists stimulated translocation of ß-arrestin2 to the cell membrane followed by ligand-receptor complex internalization without associated ß-arrestin2. In conclusion, cAMP signaling by constitutively active mutant and wild-type PTH1Rcs is inhibited by a receptor interaction with ß-arrestin2 on the cell membrane, possibly leading to uncoupling from G_s. This phenomenon is independent from protein kinases activity and the receptor C-terminal cytoplasmic domain. In addition, there are differences in the cellular localization and internalization features of constitutively active PTH1Rc mutants H223R and T410P.

This article has been cited by other articles:

				S. L. Ferrari and M. L. Bouxsein {beta}-Arrestin-Biased Parathyroid Hormone Ligands: A New Approach to the Development of Agents that Stimulate Bone Formation Science Translational Medicine, October 7, 2009; 1(1): 1ps1 - 1ps1. [Full Text] [PDF]

				C. B. Kessler and A. M. Delany Increased Notch 1 Expression and Attenuated Stimulatory G Protein Coupling to Adenylyl Cyclase in Osteonectin-Null Osteoblasts Endocrinology, April 1, 2007; 148(4): 1666 - 1674. [Abstract] [Full Text] [PDF]

				A. Rey, D. Manen, R. Rizzoli, J. Caverzasio, and S. L. Ferrari Proline-rich Motifs in the Parathyroid Hormone (PTH)/PTH-related Protein Receptor C Terminus Mediate Scaffolding of c-Src with beta-Arrestin2 for ERK1/2 Activation J. Biol. Chem., December 15, 2006; 281(50): 38181 - 38188. [Abstract] [Full Text] [PDF]

				C. A. Syme, L. Zhang, and A. Bisello Caveolin-1 Regulates Cellular Trafficking and Function of the Glucagon-Like Peptide 1 Receptor Mol. Endocrinol., December 1, 2006; 20(12): 3400 - 3411. [Abstract] [Full Text] [PDF]

				D. Gesty-Palmer, M. Chen, E. Reiter, S. Ahn, C. D. Nelson, S. Wang, A. E. Eckhardt, C. L. Cowan, R. F. Spurney, L. M. Luttrell, et al. Distinct beta-Arrestin- and G Protein-dependent Pathways for Parathyroid Hormone Receptor-stimulated ERK1/2 Activation J. Biol. Chem., April 21, 2006; 281(16): 10856 - 10864. [Abstract] [Full Text] [PDF]

				L. Wang, C. G. Radu, L. V. Yang, L. A. Bentolila, M. Riedinger, and O. N. Witte Lysophosphatidylcholine-induced Surface Redistribution Regulates Signaling of the Murine G Protein-coupled Receptor G2A Mol. Biol. Cell, May 1, 2005; 16(5): 2234 - 2247. [Abstract] [Full Text] [PDF]

				S. L. Ferrari, D. D. Pierroz, V. Glatt, D. S. Goddard, E. N. Bianchi, F. T. Lin, D. Manen, and M. L. Bouxsein Bone Response to Intermittent Parathyroid Hormone Is Altered in Mice Null for {beta}-Arrestin2 Endocrinology, April 1, 2005; 146(4): 1854 - 1862. [Abstract] [Full Text] [PDF]

				C. A. Syme, P. A. Friedman, and A. Bisello Parathyroid Hormone Receptor Trafficking Contributes to the Activation of Extracellular Signal-regulated Kinases but Is Not Required for Regulation of cAMP Signaling J. Biol. Chem., March 25, 2005; 280(12): 11281 - 11288. [Abstract] [Full Text] [PDF]

				R. Jorgensen, L. Martini, T. W. Schwartz, and C. E. Elling Characterization of Glucagon-Like Peptide-1 Receptor {beta}-Arrestin 2 Interaction: A High-Affinity Receptor Phenotype Mol. Endocrinol., March 1, 2005; 19(3): 812 - 823. [Abstract] [Full Text] [PDF]

				T. M. Murray, L. G. Rao, P. Divieti, and F. R. Bringhurst Parathyroid Hormone Secretion and Action: Evidence for Discrete Receptors for the Carboxyl-Terminal Region and Related Biological Actions of Carboxyl- Terminal Ligands Endocr. Rev., February 1, 2005; 26(1): 78 - 113. [Abstract] [Full Text] [PDF]

				A. Bisello, D. Manen, D. D. Pierroz, T. B. Usdin, R. Rizzoli, and S. L. Ferrari Agonist-Specific Regulation of Parathyroid Hormone (PTH) Receptor Type 2 Activity: Structural and Functional Analysis of PTH- and Tuberoinfundibular Peptide (TIP) 39-Stimulated Desensitization and Internalization Mol. Endocrinol., June 1, 2004; 18(6): 1486 - 1498. [Abstract] [Full Text] [PDF]

				W. B. Sneddon, C. E. Magyar, G. E. Willick, C. A. Syme, F. Galbiati, A. Bisello, and P. A. Friedman Ligand-Selective Dissociation of Activation and Internalization of the Parathyroid Hormone (PTH) Receptor: Conditional Efficacy of PTH Peptide Fragments Endocrinology, June 1, 2004; 145(6): 2815 - 2823. [Abstract] [Full Text] [PDF]

				W. B. Sneddon, C. A. Syme, A. Bisello, C. E. Magyar, M. D. Rochdi, J.-L. Parent, E. J. Weinman, A. B. Abou-Samra, and P. A. Friedman Activation-independent Parathyroid Hormone Receptor Internalization Is Regulated by NHERF1 (EBP50) J. Biol. Chem., October 31, 2003; 278(44): 43787 - 43796. [Abstract] [Full Text] [PDF]

				M. Waldhoer, P. Casarosa, M. M. Rosenkilde, M. J. Smit, R. Leurs, J. L. Whistler, and T. W. Schwartz The Carboxyl Terminus of Human Cytomegalovirus-encoded 7 Transmembrane Receptor US28 Camouflages Agonism by Mediating Constitutive Endocytosis J. Biol. Chem., May 23, 2003; 278(21): 19473 - 19482. [Abstract] [Full Text] [PDF]

				S. Chauvin, M. Bencsik, T. Bambino, and R. A. Nissenson Parathyroid Hormone Receptor Recycling: Role of Receptor Dephosphorylation and {beta}-Arrestin Mol. Endocrinol., December 1, 2002; 16(12): 2720 - 2732. [Abstract] [Full Text] [PDF]

				A. Bisello, M. Chorev, M. Rosenblatt, L. Monticelli, D. F. Mierke, and S. L. Ferrari Selective Ligand-induced Stabilization of Active and Desensitized Parathyroid Hormone Type 1 Receptor Conformations J. Biol. Chem., October 4, 2002; 277(41): 38524 - 38530. [Abstract] [Full Text] [PDF]

				M. Castro, F. Dicker, J.-P. Vilardaga, C. Krasel, M. Bernhardt, and M. J. Lohse Dual Regulation of the Parathyroid Hormone (PTH)/PTH-Related Peptide Receptor Signaling by Protein Kinase C and {beta}-Arrestins Endocrinology, October 1, 2002; 143(10): 3854 - 3865. [Abstract] [Full Text] [PDF]

				J.-P. Vilardaga, C. Krasel, S. Chauvin, T. Bambino, M. J. Lohse, and R. A. Nissenson Internalization Determinants of the Parathyroid Hormone Receptor Differentially Regulate beta -Arrestin/Receptor Association J. Biol. Chem., March 1, 2002; 277(10): 8121 - 8129. [Abstract] [Full Text] [PDF]

				S. Miserey-Lenkei, C. Parnot, S. Bardin, P. Corvol, and E. Clauser Constitutive Internalization of Constitutively Active Angiotensin II AT1A Receptor Mutants Is Blocked by Inverse Agonists J. Biol. Chem., February 15, 2002; 277(8): 5891 - 5901. [Abstract] [Full Text] [PDF]

				R. C. Gensure, P. H. Carter, B. D. Petroni, H. Juppner, and T. J. Gardella Identification of Determinants of Inverse Agonism in a Constitutively Active Parathyroid Hormone/Parathyroid Hormone-related Peptide Receptor by Photoaffinity Cross-linking and Mutational Analysis J. Biol. Chem., November 9, 2001; 276(46): 42692 - 42699. [Abstract] [Full Text] [PDF]