EHD1—An EH-Domain-Containing Protein with a Specific Expression Pattern

Mintz, Liat; Galperin, Emilia; Pasmanik‐Chor, Metsada; Tulzinsky, Sandra; Bromberg, Yael; Kozak, Christine A.; Joyner, Alexandra L.; Fein, Amos; Horowitz, Mia

doi:10.1006/geno.1999.5800

Cited by 82 publications

(136 citation statements)

References 45 publications

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“…1 A). Two immunoreactive species were observed of apparent sizes of 68 kDa (major) and 65 kDa (minor), corresponding to the sizes of Pincher (Shao et al, 2002) and the highly homologous Pincher family members, rEHD1/2 (Mintz et al, 1999;Guilherme et al, 2004), respectively. The 68 kDa protein was positively identified as Pincher in SCG neurons overexpressing an HA-tagged Pincher cDNA ( Fig.…”

Section: Pincher Mediates Trka Internalization and Endosomal Erk5 Sigmentioning

confidence: 99%

Pincher-Mediated Macroendocytosis Underlies Retrograde Signaling by Neurotrophin Receptors

Valdez¹,

Akmentin²,

Philippidou³

et al. 2005

J. Neurosci.

130

145

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Retrograde signaling by neurotrophins is crucial for regulating neuronal phenotype and survival. The mechanism responsible for retrograde signaling has been elusive, because the molecular entities that propagate Trk receptor tyrosine kinase signals from the nerve terminal to the soma have not been defined. Here, we show that the membrane trafficking protein Pincher defines the primary pathway responsible for neurotrophin retrograde signaling in neurons. By both immunofluorescence confocal and immunoelectron microscopy, we find that Pincher mediates the formation of newly identified clathrin-independent macroendosomes for Trk receptors in soma, axons, and dendrites. Trk macroendosomes are derived from plasma membrane ruffles and subsequently processed to multivesicular bodies. Pincher similarly mediates macroendocytosis for NGF (TrkA) and BDNF (TrkB) in both peripheral (sympathetic) and central (hippocampal) neurons. A unique feature of Pincher-Trk endosomes is refractoriness to lysosomal degradation, which ensures persistent signaling through a critical effector of retrograde survival signaling, Erk5 (extracellular signal-regulated kinase 5). Using sympathetic neurons grown in chamber cultures, we find that block of Pincher function, which prevents Trk macroendosome formation, eliminates retrogradely signaled neuronal survival. Pincher is the first distinguishing molecular component of a novel mechanistic pathway for endosomal signaling in neurons.

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Section: Pincher Mediates Trka Internalization and Endosomal Erk5 Sigmentioning

confidence: 99%

Pincher-Mediated Macroendocytosis Underlies Retrograde Signaling by Neurotrophin Receptors

Valdez¹,

Akmentin²,

Philippidou³

et al. 2005

J. Neurosci.

130

145

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“…26). Unlike other mammalian EH domain-containing proteins, this subgroup of four homologous proteins has a single EH domain at its C terminus (27)(28)(29)(30), a central coiled-coil region involved in oligomerization (31)(32)(33), and an N-terminal regulatory region that binds to nucleotides (32,33). Emerging evidence has implicated all four C-terminal EHD proteins in the process of endocytosis (26).…”

mentioning

confidence: 99%

EHD1 and Eps15 Interact with Phosphatidylinositols via Their Eps15 Homology Domains

Naslavsky

Rahajeng

Chenavas

et al. 2007

Journal of Biological Chemistry

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The C-terminal Eps15 homology domain-containing protein, EHD1, regulates the recycling of receptors from the endocytic recycling compartment to the plasma membrane. In cells, EHD1 localizes to tubular and spherical recycling endosomes. To date, the mode by which EHD1 associates with endosomal membranes remains unknown, and it has not been determined whether this interaction is direct or via interacting proteins. Here, we provide evidence demonstrating that EHD1 has the ability to bind directly and preferentially to an array of phospholipids, preferring phosphatidylinositols with a phosphate at position 3. Previous studies have demonstrated that EH domains coordinate calcium binding and interact with proteins containing the tripeptide asparagine-proline-phenylalanine (NPF). Using two-dimensional nuclear magnetic resonance analysis, we now describe a new function for the Eps15 homology (EH) domain of EHD1 and show that it is capable of directly binding phosphatidylinositol moieties. Moreover, we have expanded our studies to include the C-terminal EH domain of EHD4 and the second of the three N-terminal EH domains of Eps15 and demonstrated that phosphatidylinositol binding may be a more general property shared by certain other EH domains. Further studies identified a positively charged lysine residue (Lys-483) localized within the third helix of the EH domain, on the opposite face of the NPF-binding pocket, as being critical for the interaction with the phosphatidylinositols.The internalization of receptors is a critical process for eukaryotic cells. Receptors can be internalized from the plasma membrane by a variety of well described mechanisms, including via clathrin-coated pits, independently of clathrin, and through caveolae (1). Once internalized, the small vesicles containing the internalized cargo fuse with early endosomes (also known as sorting endosomes), and the receptors are then either sent to late endosomes and on to the lysosomal pathway for degradation or recycled back to the plasma membrane, where they may participate in additional rounds of internalization (2). Receptor recycling occurs either directly from the sorting endosomes in a process known as "fast recycling" or indirectly in a process termed "slow" or "regulated" recycling (3). Slow recycling has been better characterized and traverses a complex series of tubular and vesicular membrane structures that emerge from the microtubule-organizing center and is collectively known as the endocytic recycling compartment (3, 4). Despite advances in recent years, the process of recycling is not as well understood as internalization.Among the key regulatory proteins that control endocytic transport and recycling are the Rab family of GTP-binding proteins (5-7). Over 60 different mammalian Rab proteins have been identified thus far, and their highly regulated GTP binding and subsequent hydrolysis lead to recruitment of a wide array of effector proteins that are important for vesicular transport and fusion processes within the endocytic pathways. For exa...

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“…Among the EH domain-containing proteins are members of the EHD subfamily [4], characterized by three structural modules: an N-terminal domain, harboring nucleotide binding motifs [5,6]; a central coiled-coil domain, involved in oligomerization; and a single C-terminal EH domain. There are four vertebrate paralogs [7,8], two plant orthologs and one gene in C. elegans [9] and Drosophila [10]. Vertebrate EHDs show a distinct tissue expression pattern [7,8,[11][12][13] and intracellular localization.…”

Section: Introductionmentioning

confidence: 99%

“…There are four vertebrate paralogs [7,8], two plant orthologs and one gene in C. elegans [9] and Drosophila [10]. Vertebrate EHDs show a distinct tissue expression pattern [7,8,[11][12][13] and intracellular localization. EHD1 was localized mainly to the endocytic recycling compartment (ERC) [7] with some localization on the plasma membrane and early endosomes [14,15], while EHD2 was localized to the plasma membrane [16,17].…”

Section: Introductionmentioning

confidence: 99%

“…Vertebrate EHDs show a distinct tissue expression pattern [7,8,[11][12][13] and intracellular localization. EHD1 was localized mainly to the endocytic recycling compartment (ERC) [7] with some localization on the plasma membrane and early endosomes [14,15], while EHD2 was localized to the plasma membrane [16,17]. EHD3 was located on early endosomes and the perinuclear tubular and vesicular structures of the recycling compartment [12,18] and EHD4 was localized to the plasma membrane, to vesicular structures [19] and to early endosomes [20].…”

Section: Introductionmentioning

confidence: 99%

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EHDS are serine phosphoproteins: EHD1 phosphorylation is enhanced by serum stimulation

Fichtman

Ravid

Rapaport

et al. 2008

Cellular and Molecular Biology Letters

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Abbreviations used: AP-2 -adaptor protein complex-2; BSA -bovine serum albumin; CHOChinese hamster ovary; cpm -counts per minute; dFCS -dialyzed FCS; DMEM -Dulbecco modified eagle medium; EHBP1-EH domain binding protein 1; FCS -fetal calf serum; FYVE -phenylalanine-tyrosine-valine-glutamate; IGF-1R -insulin-like growth factor-1 receptor; mDHFR -mouse dihydrofolate reductase; MEM-Alpha -minimal essential medium-alpha; ORF -open reading frame; PBS -phosphate buffered saline; PKC -protein kinase C; PVDFpolyvinidilen difluoride; TLC -thin-layer chromatography; TLE -thin-layer electrophoresis Abstract: Endocytic processes are mediated by multiple protein-protein interacting modules and regulated by phosphorylation and dephosphorylation. The Eps15 homology domain containing protein 1 (EHD1) has been implicated in regulating recycling of proteins, internalized both in clathrin-dependent and clathrin-independent endocytic pathways, from the recycling compartment to the plasma membrane. EHD1 was found in a complex with clathrin, adaptor protein complex-2 (AP-2) and insulin-like growth factor-1 receptor (IGF-1R), and was shown to interact with Rabenosyn-5, SNAP29, EHBP1 (EH domain binding protein 1) and syndapin I and II. In this study, we show that EHD1, like the other human EHDs, undergoes serine-phosphorylation. Our results also indicate that EHD1 is a serum-inducible serine-phosphoprotein and that PKC (protein kinase C) is one of its kinases. In addition, we show that inhibitors of clathrin-mediated endocytosis decrease EHD1 phosphorylation, while inhibitors of caveolinmediated endocytosis do not affect EHD1 phosphorylation. The results of experiments in which inhibitors of endocytosis were employed strongly suggest that EHD1 phosphorylation occurs between early endosomes and the endocytic recycling compartment.

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EHD1—An EH-Domain-Containing Protein with a Specific Expression Pattern

Cited by 82 publications

References 45 publications

Pincher-Mediated Macroendocytosis Underlies Retrograde Signaling by Neurotrophin Receptors

Pincher-Mediated Macroendocytosis Underlies Retrograde Signaling by Neurotrophin Receptors

EHD1 and Eps15 Interact with Phosphatidylinositols via Their Eps15 Homology Domains

EHDS are serine phosphoproteins: EHD1 phosphorylation is enhanced by serum stimulation

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