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J. Biol. Chem., Vol. 279, Issue 11, 10593-10605, March 12, 2004

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EHD2 and the Novel EH Domain Binding Protein EHBP1 Couple Endocytosis to the Actin Cytoskeleton^*

Adilson Guilherme, Neil A. Soriano, Sahana Bose, John Holik, Avirup Bose, Darcy P. Pomerleau, Paul Furcinitti, John Leszyk, Silvia Corvera, and Michael P. Czech

From the Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, Massachusetts 01605

Here we identified two novel proteins denoted EH domain protein 2 (EHD2) and EHD2-binding protein 1 (EHBP1) that link clathrin-mediated endocytosis to the actin cytoskeleton. EHD2 contains an N-terminal P-loop and a C-terminal EH domain that interacts with NPF repeats in EHBP1. Disruption of EHD2 or EHBP1 function by small interfering RNA-mediated gene silencing inhibits endocytosis of transferrin into EEA1-positive endosomes as well as GLUT4 endocytosis into cultured adipocytes. EHD2 localizes with cortical actin filaments, whereas EHBP1 contains a putative actin-binding calponin homology domain. High expression of EHD2 or EHBP1 in intact cells mediates extensive actin reorganization. Thus EHD2 appears to connect endocytosis to the actin cytoskeleton through interactions of its N-terminal domain with membranes and its C-terminal EH domain with the novel EHBP1 protein.

Received for publication, July 16, 2003 , and in revised form, November 13, 2003.

^* This work was supported by National Institutes of Health Grants DK 30898 (to M. P. C.) and DK54479 (to S. C.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains QuickTime movies 1 and 2.

To whom correspondence should be addressed: Program in Molecular Medicine, University of Massachusetts Medical School, 373 Plantation St., Worcester, MA 01605. Tel.: 508-856-2254; Fax: 508-856-1617; E-mail: Michael.Czech{at}umassmed.edu.

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