TY - JOUR
T1 - Decoupling of activation and effector binding underlies ARF6 priming of fast endocytic recycling
AU - Montagnac, Guillaume
AU - De Forges, Hélène
AU - Smythe, Elizabeth
AU - Gueudry, Charles
AU - Romao, Maryse
AU - Salamero, Jean
AU - Chavrier, Philippe
N1 - Funding Information:
We gratefully acknowledge the Nikon Imaging Centre at the Institut Curie-CNRS and the PICT-IBiSA imaging facility of the Institut Curie for help with image acquisition and processing. We are indebted to G. Raposo (Curie Institute, Paris) for help with transmission electron microscopy. We thank T. Kirchhausen, M. Franco, A. Benmerah, C. D'Souza-Schorey, J.H. Keen, and M.A. Schwartz for providing reagents. Support was provided to P.C. by the Association pour la Recherche contre le Cancer, SL220100601356, and the Agence Nationale pour la Recherche, ANR-08-BLAN-0111. Core funding for this work was provided by Institut Curie and CNRS.
PY - 2011/4/12
Y1 - 2011/4/12
N2 - The small GTP-binding protein ADP-ribosylation factor 6 (ARF6) controls the endocytic recycling pathway of several plasma membrane receptors. We analyzed the localization and GDP/GTP cycle of GFP-tagged ARF6 by total internal reflection fluorescent microscopy. We found that ARF6-GFP associates with clathrin-coated pits (CCPs) at the plasma membrane in a GTP-dependent manner in a mechanism requiring the adaptor protein complex AP-2. In CCP, GTP-ARF6 mediates the recruitment of the ARF-binding domain of downstream effectors including JNK-interacting proteins 3 and 4 (JIP3 and JIP4) after the burst recruitment of the clathrin uncoating component auxilin. ARF6 does not contribute to receptor-mediated clathrin-dependent endocytosis. In contrast, we found that interaction of ARF6 and JIPs on endocytic vesicles is required for trafficking of the transferrin receptor in the fast, microtubule-dependent endocytic recycling pathway. Our findings unravel a novel mechanism of separation of ARF6 activation and effector function, ensuring that fast recycling may be determined at the level of receptor incorporation into CCPs.
AB - The small GTP-binding protein ADP-ribosylation factor 6 (ARF6) controls the endocytic recycling pathway of several plasma membrane receptors. We analyzed the localization and GDP/GTP cycle of GFP-tagged ARF6 by total internal reflection fluorescent microscopy. We found that ARF6-GFP associates with clathrin-coated pits (CCPs) at the plasma membrane in a GTP-dependent manner in a mechanism requiring the adaptor protein complex AP-2. In CCP, GTP-ARF6 mediates the recruitment of the ARF-binding domain of downstream effectors including JNK-interacting proteins 3 and 4 (JIP3 and JIP4) after the burst recruitment of the clathrin uncoating component auxilin. ARF6 does not contribute to receptor-mediated clathrin-dependent endocytosis. In contrast, we found that interaction of ARF6 and JIPs on endocytic vesicles is required for trafficking of the transferrin receptor in the fast, microtubule-dependent endocytic recycling pathway. Our findings unravel a novel mechanism of separation of ARF6 activation and effector function, ensuring that fast recycling may be determined at the level of receptor incorporation into CCPs.
UR - http://www.scopus.com/inward/record.url?scp=79953777161&partnerID=8YFLogxK
U2 - 10.1016/j.cub.2011.02.034
DO - 10.1016/j.cub.2011.02.034
M3 - Article
C2 - 21439824
AN - SCOPUS:79953777161
SN - 0960-9822
VL - 21
SP - 574
EP - 579
JO - Current Biology
JF - Current Biology
IS - 7
ER -