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First published online 12 September 2006
doi: 10.1242/jcs.03190

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Selective membrane exclusion in phagocytic and macropinocytic cups

Valentina Mercanti^1,*, Steve J. Charette^1,*, Nelly Bennett², Jean-Jeacques Ryckewaert³, François Letourneur⁴ and Pierre Cosson^1,

¹ Université de Genève, Centre Médical Universitaire, Département de Physiologie Cellulaire et Métabolisme, 1 rue Michel Servet, CH-1211 Genève 4, Switzerland
² Laboratoire de Biochimie et Biophysique des Systèmes Intégrés, Département de Réponse et Dynamique Cellulaires, CEA-Grenoble, 17 rue des Martyrs, 38054 Grenoble Cedex 9, France
³ Laboratoire de Chimie des Protéines, ERM 201 INSERM/CEA/UJF, 17 rue des Martyrs, 38054 Grenoble Cedex 9, France
⁴ Institut de Biologie et Chimie des Protéines (IBCP UMR 5086), CNRS Université de Lyon, IFR128 BioSciences Lyon-Gerland, 7 passage du Vercors, 69367 Lyon Cedex 07, France

Author for correspondence (e-mail: pierre.cosson{at}medecine.unige.ch)

Accepted 25 July 2006

Specialized eukaryotic cells can ingest large particles and sequester them within membrane-delimited phagosomes. Many studies have described the delivery of lysosomal proteins to the phagosome, but little is known about membrane sorting during the early stages of phagosome formation. Here we used Dictyostelium discoideum amoebae to analyze the membrane composition of newly formed phagosomes. The membrane delimiting the closing phagocytic cup was essentially derived from the plasma membrane, but a subgroup of proteins was specifically excluded. Interestingly the same phenomenon was observed during the formation of macropinosomes, suggesting that the same sorting mechanisms are at play during phagocytosis and macropinocytosis. Analysis of mutant strains revealed that clathrin-associated adaptor complexes AP-1, -2 and -3 were not necessary for this selective exclusion and, accordingly, ultrastructural analysis revealed no evidence for vesicular transport around phagocytic cups. Our results suggest the existence of a new, as yet uncharacterized, sorting mechanism in phagocytic and macropinocytic cups.

Key words: Membrane sorting, Phagocytosis, Macropinocytosis, Clathrin, Dictyostelium discoideum

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