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Vol. 15, Issue 3, 1134-1145, March 2004

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EFA6, Exchange Factor for ARF6, Regulates the Actin Cytoskeleton and Associated Tight Junction in Response to E-Cadherin Engagement

Frédéric Luton ^* , Stéphanie Klein ^*, Jean-Paul Chauvin , André Le Bivic , Sylvain Bourgoin , Michel Franco ^*, and Pierre Chardin ^*

^* Institut de Pharmacologie Moléculaire et Cellulaire, Centre National de la Recherche Scientifique-Unité Mixte Recherche 6097, 06560 Valbonne, France; Institut de Biologie du Développement de Marseille, Centre National de la Recherche Scientifique-Unité Mixte Recherche 6156, Faculté des Sciences de Luminy, 13288 Marseille cedex 09, France; and Centre de Recherche en Rhumatologie et Immunologie, Faculté de Médecine, Université Laval, G1V 4G2 Québec, Canada

Submitted October 21, 2003; Revised November 21, 2003; Accepted November 21, 2003
Monitoring Editor: Daniel Goodenough

We addressed the role of EFA6, exchange factor for ARF6, during the development of epithelial cell polarity in Madin-Darby canine kidney cells. EFA6 is located primarily at the apical pole of polarized cells, including the plasma membrane. After calcium-triggered E-cadherin–mediated cell adhesion, EFA6 is recruited to a Triton X-100–insoluble fraction and its protein level is increased concomitantly to the accelerated formation of a functional tight junction (TJ). The expression of EFA6 results in the selective retention at the cell surface of the TJ protein occludin. This effect is due to EFA6 capacities to promote selectively the stability of the apical actin ring onto which the TJ is anchored, resulting in the exclusion of TJ proteins from endocytosis. Finally, our data suggest that EFA6 effects are achieved by the coordinate action of both its exchange activity and its actin remodeling C-terminal domain. We conclude that EFA6 is a signaling molecule that responds to E-cadherin engagement and is involved in TJ formation and stability.

Abbreviations used: AJ, adherens junction; Dox, doxycycline; GAP, GTPase activating protein; GEF, guanine nucleotide exchange factor; IF, immunofluorescence; PH, pleckstrin homology; PM, plasma membrane; RITC, rhodamine B isothiocyanate; TER, transepithelial resistance; Tf-R, transferrin receptor, TJ, tight junction.

Corresponding author. E-mail address: luton{at}ipmc.cnrs.fr.

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