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Published 8 December 2003. doi:10.1083/jcb.200305029
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© The Rockefeller University Press, 0021-9525/2003/12/1111 $8.00
The Journal of Cell Biology, Volume 163, Number 5, 1111-1121

Article

ARF6 controls post-endocytic recycling through its downstream exocyst complex effector



Magali Prigent1, Thierry Dubois1, Graça Raposo2, Valérie Derrien1, Danièle Tenza2, Carine Rossé3, Jacques Camonis3 and Philippe Chavrier1

1 Membrane and Cytoskeleton Dynamics Group, Centre National de la Recherche Scientifique, UMR-144
2 Electron Microscopy Laboratory, Centre National de la Recherche Scientifique, UMR-144
3 Institut National de la Santé et de la Recherche Médicale, U-528, Institut Curie, F-75248 Paris Cedex 05, France

Address correspondence to Philippe Chavrier, UMR144 Centre National de la Recherche Scientifique, Institut Curie, 26 rue d'Ulm, F-75248 Paris cedex 05, France. Tel.: 33-1-42-34-63-59. Fax: 33-1-42-34-63-77. email: philippe.chavrier{at}curie.fr

The small guanosine triphosphate (GTP)–binding protein ADP-ribosylation factor (ARF) 6 regulates membrane recycling to regions of plasma membrane remodeling via the endocytic pathway. Here, we show that GTP–bound ARF6 interacts with Sec10, a subunit of the exocyst complex involved in docking of vesicles with the plasma membrane. We found that Sec10 localization in the perinuclear region is not restricted to the trans-Golgi network, but extends to recycling endosomes. In addition, we report that depletion of Sec5 exocyst subunit or dominant inhibition of Sec10 affects the function and the morphology of the recycling pathway. Sec10 is found to redistribute to ruffling areas of the plasma membrane in cells expressing GTP-ARF6, whereas dominant inhibition of Sec10 interferes with ARF6-induced cell spreading. Our paper suggests that ARF6 specifies delivery and insertion of recycling membranes to regions of dynamic reorganization of the plasma membrane through interaction with the vesicle-tethering exocyst complex.

Key Words: ARF6; exocyst complex; recycling; endocytosis; small GTP-binding protein

The online version of this article contains supplemental material.

Abbreviations used in this paper: ARF, ADP-ribosylation factor; NRK, normal rat kidney; RE, recycling endosome; siRNA, small interfering RNA; Tf, transferrin; TfR, transferrin-receptor.


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