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J. Biol. Chem., Vol. 279, Issue 34, 35687-35691, August 20, 2004

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Crucial Role of Neuron-enriched Endosomal Protein of 21 kDa in Sorting between Degradation and Recycling of Internalized G-protein-coupled Receptors^*

Colin Debaigt, Harald Hirling, Pascal Steiner, Jean-Pierre Vincent, and Jean Mazella¶

From the Institut de Pharmacologie Moléculaire et Cellulaire, Unité Mixte de Recherche 6097 du Centre National de la Recherche Scientifique, 660 route des Lucioles, Sophia Antipolis, 06560 Valbonne, France and Faculté des Sciences de la Vie, Ecole Polytechnique Fédérale de Lausanne, 1015 Lausanne, Switzerland

Recycling of endocytosed G-protein-coupled receptors involves a series of molecular events through early and recycling endosomes. The purpose of this work was to study the role of neuron-enriched endosomal protein of 21 kDa (NEEP21) in the recycling process of neurotensin receptors-1 and -2. Here we showed that suppression of NEEP21 expression does not modify the internalization rate of both receptors but strongly inhibited the recycling of the neurotensin receptor-2. In contrast, overexpression of NEEP21 changes the behavior of the neurotensin receptor-1 from a non-recycling to a recycling state. Recycling of the neurotensin receptor-2 involves both the phosphatidylinositol 3-kinase and the recycling endosome pathways, whereas recycling of the neurotensin receptor-1 induced by overexpression of NEEP21 only occurs by the phosphatidylinositol 3-kinase-dependent pathway. Taken together, these results confirm the essential role of NEEP21 in the recycling mechanism and show that this protein acts at the level of early endosomes to promote sorting of receptors toward a recycling pathway.

Received for publication, March 11, 2004 , and in revised form, June 3, 2004.

^* This study was supported by Grant PICS number 2051 (to J. M.) from the Centre National de la Recherche Scientifique (CNRS) and by Grants 3100AO-100834/1 from the Swiss National Science Foundation and 1907/ep from the Leenaards Foundation (to H. H.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^¶ To whom correspondence should be addressed. Tel.: 33-4-93-95-77-61; Fax: 33-4-93-95-77-08; E-mail: mazella{at}ipmc.cnrs.fr.

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