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, Steven H Caplan

Research output: Contribution to journalArticle

52 Citations (Scopus)

Abstract

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
Volume20
Issue number11
DOIs
StatePublished - Jun 1 2009

Fingerprint

Recycling
Phosphatidylinositols
Membranes
Small Interfering RNA
Cell Membrane
Transferrin
Endocytosis
Integrins
Transfection
Proteins
Magnetic Resonance Spectroscopy
phosphatidylinositol 4-phosphate
Lipids

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

@article{93dafd518b174b81ae88d15b623e495d,
title = "Eps15 homology domain 1-associated tubules contain phosphatidylinositol-4- phosphate and phosphatidylinositol-(4,5)-bisphosphate and are required for efficient recycling",
abstract = "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.",
author = "Marko Jović and Fabien Kieken and Naava Naslavsky and Sorgen, {Paul L} and Caplan, {Steven H}",
year = "2009",
month = "6",
day = "1",
doi = "10.1091/mbc.E08-11-1102",
language = "English (US)",
volume = "20",
pages = "2731--2743",
journal = "Molecular Biology of the Cell",
issn = "1059-1524",
publisher = "American Society for Cell Biology",
number = "11",

}

TY - JOUR

T1 - Eps15 homology domain 1-associated tubules contain phosphatidylinositol-4- phosphate and phosphatidylinositol-(4,5)-bisphosphate and are required for efficient recycling

AU - Jović, Marko

AU - Kieken, Fabien

AU - Naslavsky, Naava

AU - Sorgen, Paul L

AU - Caplan, Steven H

PY - 2009/6/1

Y1 - 2009/6/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=66349123530&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=66349123530&partnerID=8YFLogxK

U2 - 10.1091/mbc.E08-11-1102

DO - 10.1091/mbc.E08-11-1102

M3 - Article

C2 - 19369419

AN - SCOPUS:66349123530

VL - 20

SP - 2731

EP - 2743

JO - Molecular Biology of the Cell

JF - Molecular Biology of the Cell

SN - 1059-1524

IS - 11

ER -