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Published online 16 June 2003. doi:10.1083/jcb.200210069
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© The Rockefeller University Press, 0021-9525/2003/6/1143 $5.00
The Journal of Cell Biology, Volume 161, Number 6, 1143-1150

Article

 ADP ribosylation factor 6 is activated and controls membrane delivery during phagocytosis in macrophages

Florence Niedergang1, Emma Colucci-Guyon1, Thierry Dubois1, Graça Raposo2 and Philippe Chavrier1

1 Membrane and Cytoskeleton Dynamics Group, UMR 144 Centre National de la Recherche Scientifique, Institut Curie, F-75248 Paris Cedex 05, France
2 Electron Microscopy Group, UMR 144 Centre National de la Recherche Scientifique, Institut Curie, F-75248 Paris Cedex 05, France

Address correspondence to Florence Niedergang, Membrane and Cytoskeleton Dynamics Laboratory, UMR144 CNRS, Institut Curie, 26 rue d'Ulm, F-75248 Paris cedex 05, France. Tel.: 33-1-42-34-63-67. Fax: 33-1-42-34-63-77. E-mail: Florence.Niedergang{at}curie.fr

Engulfment of particles by phagocytes is induced by their interaction with specific receptors on the cell surface, which leads to actin polymerization and the extension of membrane protrusions to form a closed phagosome. Membrane delivery from internal pools is considered to play an important role in pseudopod extension during phagocytosis. Here, we report that endogenous ADP ribosylation factor 6 (ARF6), a small GTP-binding protein, undergoes a sharp and transient activation in macrophages when phagocytosis was initiated via receptors for the Fc portion of immunoglobulins (FcRs). A dominant-negative mutant of ARF6 (T27N mutation) dramatically affected FcR-mediated phagocytosis. Expression of ARF6-T27N lead to a reduction in the focal delivery of vesicle-associated membrane protein 3+ endosomal recycling membranes at phagocytosis sites, whereas actin polymerization was unimpaired. This resulted in an early blockade in pseudopod extension and accumulation of intracellular vesicles, as observed by electron microscopy. We conclude that ARF6 is a major regulator of membrane recycling during phagocytosis.

Key Words: VAMP3/cellubrevin; ARF6; recycling; endocytosis; actin

* Abbreviations used in this paper: ARF, ADP ribosylation factor; FcR, receptor for the Fc portion of immunoglobulins; GGA, Golgi-localized {gamma} ear–containing ARF-binding protein; PI3K, phosphatidylinositol 3-kinase; SRBC, sheep red blood cell; Tf, transferrin; VAMP, vesicle-associated membrane protein; WT, wild type.


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