Eps15 homology domain 1-associated tubules contain phosphatidylinositol-4- phosphate and phosphatidylinositol-(4,5)-bisphosphate and are required for efficient recycling

Marko Jović, Fabien Kieken, Naava Naslavsky, Paul L. Sorgen, Steve Caplan

Research output: Contribution to journalArticle

53 Scopus citations


The C-terminal Eps15 homology domain (EHD) 1/receptor-mediated endocytosis-1 protein regulates recycling of proteins and lipids from the recycling compartment to the plasma membrane. Recent studies have provided insight into the mode by which EHD1-associated tubular membranes are generated and the mechanisms by which EHD1 functions. Despite these advances, the physiological function of these striking EHD1-associated tubular membranes remains unknown. Nuclear magnetic resonance spectroscopy demonstrated that the Eps15 homology (EH) domain of EHD1 binds to phosphoinositides, including phosphatidylinositol-4-phosphate. Herein, we identify phosphatidylinositol-4- phosphate as an essential component of EHD1-associated tubules in vivo. Indeed, an EHD1 EH domain mutant (K483E) that associates exclusively with punctate membranes displayed decreased binding to phosphatidylinositol-4-phosphate and other phosphoinositides. Moreover, we provide evidence that although the tubular membranes to which EHD1 associates may be stabilized and/or enhanced by EHD1 expression, these membranes are, at least in part, pre-existing structures. Finally, to underscore the function of EHD1-containing tubules in vivo, we used a small interfering RNA (siRNA)/rescue assay. On transfection, wild-type, tubule-associated, siRNA-resistant EHD1 rescued transferrin and β1 integrin recycling defects observed in EHD1-depleted cells, whereas expression of the EHD1 K483E mutant did not. We propose that phosphatidylinositol-4-phosphate is an essential component of EHD1-associated tubules that also contain phosphatidylinositol-( 4,5)-bisphosphate and that these structures are required for efficient recycling to the plasma membrane.

Original languageEnglish (US)
Pages (from-to)2731-2743
Number of pages13
JournalMolecular biology of the cell
Issue number11
Publication statusPublished - Jun 1 2009


ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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