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Vol. 15, Issue 8, 3698-3708, August 2004

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The Cell Fate Determinant Numb Interacts with EHD/Rme-1 Family Proteins and Has a Role in Endocytic Recycling

Christian A. Smith ^* , Sascha E. Dho ^*, Julie Donaldson , Ulrich Tepass , and C. Jane McGlade ^*  ||

^* Arthur and Sonia Labatt Brain Tumour Research Centre, Hospital for Sick Children, Toronto, Ontario, Canada, M5G 1X8; Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada M5G 2M9; Laboratory of Cell Biology, National Heart Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892; and Department of Zoology, University of Toronto, Toronto, Ontario, Canada M5S 3G5

Submitted January 14, 2004; Revised May 5, 2004; Accepted May 11, 2004
Monitoring Editor: Jennifer Lippincott-Schwartz

The adaptor protein Numb is necessary for the cell fate specification of progenitor cells in the Drosophila nervous system. Numb is evolutionarily conserved and previous studies have provided evidence for a similar functional role during mammalian development. The Numb protein has multiple protein-protein interaction regions including a phosphotyrosine binding (PTB) domain and a carboxy-terminal domain that contains conserved interaction motifs including an EH (Eps15 Homology) domain binding motif and -adaptin binding site. In this study we identify the EHD/Rme-1/Pincher family of endocytic proteins as Numb interacting partners in mammals and Drosophila. The EHD/Rme-1 proteins function in recycling of plasma membrane receptors internalized by both clathrin-mediated endocytosis and a clathrin-independent pathway regulated by ADP ribosylation factor 6 (Arf6). Here we report that Numb colocalizes with endogenous EHD4/Pincher and Arf6 and that Arf6 mutants alter Numb subcellular localization. In addition, we present evidence that Numb has a novel function in endosomal recycling and intracellular trafficking of receptors.

^|| Corresponding author. E-mail address: jmcglade{at}sickkids.ca.

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