An actin-binding protein of the Sla2/Huntingtin interacting protein 1 family is a novel component of clathrin-coated pits and vesicles.
AuthorsEngqvist-Goldstein AE, Kessels MM, Chopra VS, Hayden MR, Drubin DG
JournalJ Cell Biol
PubMed ID10613908
'The actin cytoskeleton has been implicated in endocytosis, yet few molecules that link these systems have been identified. Here, we have cloned and characterized mHip1R, a protein that is closely related to huntingtin interacting protein 1 (Hip1). These two proteins are mammalian homologues of Sla2p, an actin binding protein important ... More
Overexpression of proteins containing tyrosine- or leucine-based sorting signals affects transferrin receptor trafficking.
AuthorsNordeng TW, Bakke O
JournalJ Biol Chem
PubMed ID10409667
'Targeting of many transmembrane proteins to post-Golgi compartments is dependent on cytoplasmically exposed sorting signals. The most widely used signals conform to the tyrosine- or the leucine-based motifs. Both types of signals have been implicated in protein localization to the same intracellular compartments, but previous results from both cell-free experiments ... More
Syntaxin 13 mediates cycling of plasma membrane proteins via tubulovesicular recycling endosomes.
AuthorsPrekeris R, Klumperman J, Chen YA, Scheller RH
JournalJ Cell Biol
PubMed ID9817754
'Endocytosis-mediated recycling of plasma membrane is a critical vesicle trafficking step important in diverse biological processes. The membrane trafficking decisions and sorting events take place in a series of heterogeneous and highly dynamic organelles, the endosomes. Syntaxin 13, a recently discovered member of the syntaxin family, has been suggested to ... More
A role for ADP-ribosylation factor 6 in the processing of G-protein-coupled receptors.
AuthorsMadziva MT, Birnbaumer M
JournalJ Biol Chem
PubMed ID16497672
'After agonist-induced internalization, the vasopressin V2 receptor (V2R) does not recycle to the plasma membrane. The ADP-ribosylation factor (ARF) proteins initiate vesicular intracellular traffic by promoting the recruitment of adaptor proteins; thus, we sought to determine whether ARF6 could promote V2R recycling. Neither the agonist-induced internalization nor the recycling of ... More
High cell sensitivity to Helicobacter pylori VacA toxin depends on a GPI-anchored protein and is not blocked by inhibition of the clathrin-mediated pathway of endocytosis.
AuthorsRicci V, Galmiche A, Doye A, Necchi V, Solcia E, Boquet P
JournalMol Biol Cell
PubMed ID11071915
'Helicobacter pylori vacuolating toxin (VacA) causes vacuolation in a variety of cultured cell lines, sensitivity to VacA differing greatly, however, among the different cell types. We found that the high sensitivity of HEp-2 cells to VacA was impaired by treating the cells with phosphatidylinositol-specific phospholipase C (PI-PLC) which removes glycosylphosphatidylinositol ... More
Bending a membrane: how clathrin affects budding.
AuthorsHinrichsen L, Meyerholz A, Groos S, Ungewickell EJ
JournalProc Natl Acad Sci U S A
PubMed ID16735469
'Receptor-mediated endocytosis of ligands, such as transferrin and LDL, is suppressed when clathrin synthesis is blocked by RNA interference in HeLa cells. We have found that domains containing the adapter complex 2 (AP2)-coated vesicle adapter and the endocytic accessory proteins CALM (clathrin assembly lymphoid myeloid leukemia protein), epsin, and eps15/eps15R ... More
Morphine activates opioid receptors without causing their rapid internalization.
AuthorsKeith DE, Murray SR, Zaki PA, Chu PC, Lissin DV, Kang L, Evans CJ, von Zastrow M
JournalJ Biol Chem
PubMed ID8702570
'We have examined the endocytic trafficking of epitope-tagged delta and mu opioid receptors expressed in human embryonic kidney (HEK) 293 cells. These receptors are activated by peptide agonists (enkephalins) as well as by the alkaloid agonist drugs etorphine and morphine. Enkephalins and etorphine cause opioid receptors to internalize rapidly (t1/2 ... More
Distribution and function of AP-1 clathrin adaptor complexes in polarized epithelial cells.
AuthorsFölsch H, Pypaert M, Schu P, Mellman I
JournalJ Cell Biol
PubMed ID11157985
'Expression of the epithelial cell-specific heterotetrameric adaptor complex AP-1B is required for the polarized distribution of many membrane proteins to the basolateral surface of LLC-PK1 kidney cells. AP-1B is distinguished from the ubiquitously expressed AP-1A by exchange of its single 50-kD mu subunit, mu1A, being replaced by the closely related ... More
Secretion, surface localization, turnover, and steady state expression of protein disulfide isomerase in rat hepatocytes.
'Protein disulfide isomerase in isolated rat hepatocytes was present at a concentration of 7 micrograms/mg cell protein, representing a approximately 2-fold enrichment compared to isolated hepatic non-parenchymal cells. Though localized mainly in microsomal fractions of hepatocytes, direct immunofluorescence and cell surface radioiodination followed by immunoprecipitation revealed the presence of M(r) ... More
The trimeric GTP-binding protein (G(q)/G(11)) alpha subunit is required for insulin-stimulated GLUT4 translocation in 3T3L1 adipocytes.
AuthorsKanzaki M, Watson RT, Artemyev NO, Pessin JE
JournalJ Biol Chem
PubMed ID10702285
'To investigate the potential role of trimeric GTP-binding proteins regulating GLUT4 translocation in adipocytes, wild type and constitutively active G(q) (G(q)/Q209L), G(i) (G(i)/Q205L), and G(s) (G(s)/Q227L) alpha subunit mutants were expressed in 3T3L1 adipocytes. Although expression of neither the wild type nor G(i)/Q205L and G(s)/Q227L alpha subunit mutants had any ... More
pH-independent retrograde targeting of glycolipids to the Golgi complex.
AuthorsSchapiro FB, Lingwood C, Furuya W, Grinstein S
JournalAm J Physiol
PubMed ID9486120
'A small fraction of the molecules internalized by endocytosis reaches the Golgi complex through a retrograde pathway that is poorly understood. In the present work, we used bacterial toxins to study the retrograde pathway in Vero cells. The recombinant B subunit of verotoxin 1B (VT1B) was labeled with fluorescein to ... More
Synaptotagmin VII regulates Ca(2+)-dependent exocytosis of lysosomes in fibroblasts.
'Synaptotagmins (Syts) are transmembrane proteins with two Ca(2+)-binding C(2) domains in their cytosolic region. Syt I, the most widely studied isoform, has been proposed to function as a Ca(2+) sensor in synaptic vesicle exocytosis. Several of the twelve known Syts are expressed primarily in brain, while a few are ubiquitous ... More
Sphingolipids and glycoproteins are differentially trafficked to the Chlamydia trachomatis inclusion.
AuthorsScidmore MA, Fischer ER, Hackstadt T
JournalJ Cell Biol
PubMed ID8707822
'Chlamydia trachomatis is an obligate intracellular pathogen that multiples within the confines of a membrane-bound vacuole called an inclusion. Approximately 40-50% of the sphingomyelin synthesized from exogenously added NBD-ceramide is specifically transported from the Golgi apparatus to the chlamydial inclusion (Hackstadt, T., M.A. Scidmore, and D.D. Rockey. 1995. Proc. Natl. ... More
GLUT4 and transferrin receptor are differentially sorted along the endocytic pathway in CHO cells.
AuthorsWei ML, Bonzelius F, Scully RM, Kelly RB, Herman GA
JournalJ Cell Biol
PubMed ID9456317
'The trafficking of GLUT4, a facilitative glucose transporter, is examined in transfected CHO cells. In previous work, we expressed GLUT4 in neuroendocrine cells and fibroblasts and found that it was targeted to a population of small vesicles slightly larger than synaptic vesicles (Herman, G.A, F. Bonzelius, A.M. Cieutat, and R.B. ... More
N-terminal protein acylation confers localization to cholesterol, sphingolipid-enriched membranes but not to lipid rafts/caveolae.
AuthorsMcCabe JB, Berthiaume LG
JournalMol Biol Cell
PubMed ID11694592
'When variably fatty acylated N-terminal amino acid sequences were appended to a green fluorescent reporter protein (GFP), chimeric GFPs were localized to different membranes in a fatty acylation-dependent manner. To explore the mechanism of localization, the properties of acceptor membranes and their interaction with acylated chimeric GFPs were analyzed in ... More
Luminescence quenching by long range electron transfer: a probe of protein clustering and conformation at the cell surface.
AuthorsMatko J, Jenei A, Wei T, Edidin M
JournalCytometry
PubMed ID7736865
'Quenching of luminescence from fluorescent and phosphorescent probes by nitroxide spin labels with a long range electron transfer (LRET) mechanism (44,45) has been tested as a tool to monitor association/clustering and conformational changes of cell surface proteins. The membrane proteins were labeled with monoclonal antibodies or Fab fragments conjugated with ... More
The tyrosine kinase ACK1 associates with clathrin-coated vesicles through a binding motif shared by arrestin and other adaptors.
AuthorsTeo M, Tan L, Lim L, Manser E
JournalJ Biol Chem
PubMed ID11278436
'One target for the small GTPase Cdc42 is the nonreceptor tyrosine kinase activated Cdc42-associated kinase (ACK), which binds selectively to Cdc42.GTP. We report that ACK1 can associate directly with the heavy chain of clathrin. A central region in ACK1 containing a conserved motif behaves as a clathrin adaptor and competes ... More
Localization of cellubrevin-related peptide, endobrevin, in the early endosome in pancreatic beta cells and its physiological function in exo-endocytosis of secretory granules.
AuthorsNagamatsu S, Nakamichi Y, Watanabe T, Matsushima S, Yamaguchi S, Ni J, Itagaki E, Ishida H
JournalJ Cell Sci
PubMed ID11112705
'Cellubrevins are integral membrane proteins expressed in a wide variety of tissues and usually localized in recycling vesicles. Here, we investigated the cellular localization of a cellubrevin-related peptide, endobrevin, in pancreatic (beta) cells and its implication in the exo-endocytosis of insulin and (gamma)-amino butyric acid (GABA). Immunocytochemistry showed that endobrevin ... More
Cytoplasmic and nuclear delivery of a TAT-derived peptide and a beta-peptide after endocytic uptake into HeLa cells.
AuthorsPotocky TB, Menon AK, Gellman SH
JournalJ Biol Chem
PubMed ID14517218
'Several short, highly cationic peptides are able to enter the cytoplasm and nucleus of cells from the extracellular medium. The mechanism of entry is unknown. A number of fluorescence-based studies suggested that these molecules cross the plasma membrane by an energy-independent process, directly gaining access to the cytoplasm. Recent reports ... More
Wortmannin alters the transferrin receptor endocytic pathway in vivo and in vitro.
AuthorsSpiro DJ, Boll W, Kirchhausen T, Wessling-Resnick M
JournalMol Biol Cell
PubMed ID8868465
'Treatment with the phosphatidylinositol 3-kinase inhibitor wortmannin promotes approximately 30% decrease in the steady-state number of cell-surface transferrin receptors. This effect is rapid and dose dependent, with maximal down-regulation elicited with 30 min of treatment and with an IC50 approximately 25 nM wortmannin. Wortmannin-treated cells display an increased endocytic rate ... More
Applications of ratio fluorescence microscopy in the study of cell physiology.
AuthorsDunn KW, Mayor S, Myers JN, Maxfield FR
JournalFASEB J
PubMed ID8005385
'Quantitative fluorescence microscopy is becoming an increasingly important tool in the study of cell biology. Fluorescence microscopy has long been used for qualitative characterizations of subcellular distributions of proteins, lipids, nucleic acids, and ions, but quantifying these distributions is complicated by a variety of optical, biological, and physical factors. Many ... More
The subcellular distribution of early endosomes is affected by the annexin II2p11(2) complex.
AuthorsHarder T, Gerke V
JournalJ Cell Biol
PubMed ID8245122
'The tyrosine kinase substrate annexin II is a member of a multigene family of Ca2+ and lipid-binding proteins which have been implicated in a number of membrane-related events. We have analyzed the subcellular distribution of annexin II in relation to other cellular components in normal and specifically manipulated MDCK cells. ... More
Formation of mutually exclusive Rab11 complexes with members of the family of Rab11-interacting proteins regulates Rab11 endocytic targeting and function.
AuthorsMeyers JM, Prekeris R
JournalJ Biol Chem
PubMed ID12376546
'Several Rabs, including Rab11, regulate the traffic and sorting of proteins in the endosomal pathway. Recently, six novel Rab11 family interacting proteins (FIPs) were identified. Although they share little overall sequence homology, all FIPs contain a conserved Rab11-binding domain. Here we investigate the role of FIPs as Rab11-targeting proteins and ... More
Movement of internalized ligand-receptor complexes along a continuous endosomal reticulum.
AuthorsHopkins CR, Gibson A, Shipman M, Miller K
JournalNature
PubMed ID2374607
'Complexes of cell-surface receptors and their ligands are commonly internalized by endocytosis and enter a prelysosomal endosomal pathway for further processing. Fluorescence microscopy and video recording of living cells to trace the passage of ligand-receptor complexes has identified the endosomal compartment as an extensive network of tubular cisternae. Endocytosed material ... More
Novel guanidine-containing molecular transporters based on lactose scaffolds: lipophilicity effect on the intracellular organellar selectivity.
AuthorsBiswas G, Jeon OY, Lee WS, Kim DC, Kim KT, Lee S, Chang S, Chung SK,
JournalChemistry
PubMed ID18770513
We have synthesized two lactose-based molecular transporters, each containing seven guanidine residues attached to the lactose scaffold through omega-aminocarboxylate linker chains of two different lengths, and have examined their cellular uptakes and intracellular and organellar localizations in HeLa cells, as well as their tissue distributions in mice. Both molecular transporters ... More
Intercellular trafficking and protein delivery by a herpesvirus structural protein.
AuthorsElliott G, O'Hare P,
JournalCell
PubMed ID9008163
We show that the HSV-1 structural protein VP22 has the remarkable property of intercellular transport, which is so efficient that following expression in a subpopulation the protein spreads to every cell in a monolayer, where it concentrates in the nucleus and binds chromatin. VP22 movement was observed both after delivery ... More
Regulation of receptor-mediated endocytosis by Rho and Rac.
Pinocytosis and membrane ruffling are among the earliest and most dramatic cellular responses to stimulation by growth factors or other mitogens. The small Ras-related G proteins Rho and Rac have a regulatory role in membrane ruffling and activated Rho has been shown to stimulate pinocytosis when microinjected into Xenopus oocytes. ... More
Actin assembly at membranes controlled by ARF6.
AuthorsSchafer DA, D'Souza-Schorey C, Cooper JA
JournalTraffic
PubMed ID11273133
The small GTPase, ADP-ribosylation factor-6 (ARF6), has been implicated in regulating membrane traffic and remodeling cortical F-actin. Using real-time video analysis of actin assembly in living cells, we investigated the function and mechanism of ARF6 in control of actin assembly. Expression of an activated form of ARF6 that mimicks the ... More
Mammalian class E vps proteins recognize ubiquitin and act in the removal of endosomal protein-ubiquitin conjugates.
AuthorsBishop N, Horman A, Woodman P
JournalJ Cell Biol
PubMed ID11916981
There is increasing evidence that ubiquitination of receptors provides an important endosomal sorting signal. Here we report that mammalian class E vacuolar protein-sorting (vps) proteins recognize ubiquitin. Both tumor susceptibility gene 101 (TSG101)/human VPS (hVPS)28 and hepatocyte growth factor receptor substrate (Hrs) cytosolic complexes bind ubiquitin-agarose. TSG101 and hVPS28 are ... More
Consequences of the selective blockage of chaperone-mediated autophagy.
AuthorsMassey AC, Kaushik S, Sovak G, Kiffin R, Cuervo AM
JournalProc Natl Acad Sci U S A
PubMed ID16585521
Chaperone-mediated autophagy (CMA) is a selective pathway for the degradation of cytosolic proteins in lysosomes. CMA declines with age because of a decrease in the levels of lysosome-associated membrane protein (LAMP) type 2A, a lysosomal receptor for this pathway. We have selectively blocked the expression of LAMP-2A in mouse fibroblasts ... More
Rab11-FIP2, an adaptor protein connecting cellular components involved in internalization and recycling of epidermal growth factor receptors.
AuthorsCullis DN, Philip B, Baleja JD, Feig LA
JournalJ Biol Chem
PubMed ID12364336
Rab11-FIP2 is a member of a newly identified family of Rab11-binding proteins that have been implicated in the function of recycling endosomes. Here we show that Rab11-FIP2 may also be involved with the process of receptor-mediated endocytosis. First we demonstrate that Rab11-FIP2 contains an NPF motif that allows it to ... More
Characterization of the cytosolic tuberin-hamartin complex. Tuberin is a cytosolic chaperone for hamartin.
AuthorsNellist M, van Slegtenhorst MA, Goedbloed M, van den Ouweland AM, Halley DJ, van der Sluijs P
JournalJ Biol Chem
PubMed ID10585443
Tuberous sclerosis (TSC) is an autosomal dominant disorder characterized by a broad phenotypic spectrum that includes seizures, mental retardation, renal dysfunction and dermatological abnormalities. Mutations to either the TSC1 or TSC2 gene are responsible for the disease. The TSC1 gene encodes hamartin, a 130-kDa protein without significant homology to other ... More
Selective inhibition of transcytosis by brefeldin A in MDCK cells.
AuthorsHunziker W, Whitney JA, Mellman I
JournalCell
PubMed ID1934063
Treatment of most cells with brefeldin A (BFA) leads to the retrieval of the Golgi complex to the endoplasmic reticulum, presumably reflecting an inhibition of cytoplasmic coat protein binding to Golgi membranes. Although BFA has been thought to act only on biosynthetic organelles, we now show that this drug also ... More
A phosphatidylinositol (4,5)-bisphosphate binding site within mu2-adaptin regulates clathrin-mediated endocytosis.
AuthorsRohde G, Wenzel D, Haucke V
JournalJ Cell Biol
PubMed ID12119359
The clathrin adaptor complex AP-2 serves to coordinate clathrin-coated pit assembly with the sorting of transmembrane cargo proteins at the plasmalemma. How precisely AP-2 assembly and cargo protein recognition at sites of endocytosis are regulated has remained unclear, but recent evidence implicates phosphoinositides, in particular phosphatidylinositol (4,5)-bisphosphate (PI[4,5]P2), in these ... More
Functional expression cloning and characterization of SFT, a stimulator of Fe transport.
AuthorsGutierrez JA, Yu J, Rivera S, Wessling-Resnick M
JournalJ Cell Biol
PubMed ID9362508
A stimulator of Fe transport (SFT) was identified by functional expression cloning in Xenopus oocytes. SFT-mediated transport has properties defined for transferrin-independent Fe uptake, but its cytolocalization in recycling endosomes and the observed stimulation of transferrin-bound Fe assimilation indicate a key role in intracellular Fe membrane transport as well. SFT ... More
Recycling of the insulin-sensitive glucose transporter GLUT4. Access of surface internalized GLUT4 molecules to the perinuclear storage compartment is mediated by the Phe5-Gln6-Gln7-Ile8 motif.
AuthorsPalacios S, Lalioti V, Martinez-Arca S, Chattopadhyay S, Sandoval IV
JournalJ Biol Chem
PubMed ID11031262
The insulin-sensitive glucose transporter GLUT4 is translocated to the plasma membrane in response to insulin and recycled back to the intracellular store(s) after removal of the hormone. We have used clonal 3T3-L1 fibroblasts and adipocyte-like cells stably expressing wild-type GLUT4 to characterize (a) the intracellular compartment where the bulk of ... More
Clathrin-independent pinocytosis is induced in cells overexpressing a temperature-sensitive mutant of dynamin.
AuthorsDamke H, Baba T, van der Bliek AM, Schmid SL
JournalJ Cell Biol
PubMed ID7559787
A stable HeLa cell line expressing a dynamin mutant, dynts, exhibits a temperature-sensitive defect in endocytic clathrin-coated vesicle formation. Dynts carries a point mutation, G273D, corresponding to the Drosophila shibirets1 allele. The ts-defect in receptor-mediated endocytosis shows a rapid onset (< 5 min) and is readily reversible. At the nonpermissive ... More
Overexpression of tau protein inhibits kinesin-dependent trafficking of vesicles, mitochondria, and endoplasmic reticulum: implications for Alzheimer's disease.
AuthorsEbneth A, Godemann R, Stamer K, Illenberger S, Trinczek B, Mandelkow E
JournalJ Cell Biol
PubMed ID9813097
The neuronal microtubule-associated protein tau plays an important role in establishing cell polarity by stabilizing axonal microtubules that serve as tracks for motor-protein-driven transport processes. To investigate the role of tau in intracellular transport, we studied the effects of tau expression in stably transfected CHO cells and differentiated neuroblastoma N2a ... More
Identification of the universal cofactor (auxilin 2) in clathrin coat dissociation.
AuthorsUmeda A, Meyerholz A, Ungewickell E
JournalEur J Cell Biol
PubMed ID10887964
Uncoating of clathrin-coated vesicles in neuronal cells requires hsc70 in concert with the cofactor auxilin which contains a J-domain as well as a domain with homology to dual specific phosphatases and tensin, known as PTEN. The question of whether an analogous factor operates in other cell types has until now ... More
Copper-stimulated endocytosis and degradation of the human copper transporter, hCtr1.
AuthorsPetris MJ, Smith K, Lee J, Thiele DJ
JournalJ Biol Chem
PubMed ID12501239
Copper uptake at the plasma membrane and subsequent delivery to copper-dependent enzymes is essential for many cellular processes, including mitochondrial oxidative phosphorylation, free radical detoxification, pigmentation, neurotransmitter synthesis, and iron metabolism. However, intracellular levels of this nutrient must be controlled because it is potentially toxic in excess concentrations. The hCtr1 ... More
Rapid endo-lysosomal escape of poly(DL-lactide-co-glycolide) nanoparticles: implications for drug and gene delivery.
AuthorsPanyam J, Zhou WZ, Prabha S, Sahoo SK, Labhasetwar V
JournalFASEB J
PubMed ID12153989
The endo-lysosomal escape of drug carriers is crucial to enhancing the efficacy of their macromolecular payload, especially the payloads that are susceptible to lysosomal degradation. Current vectors that enable the endo-lysosomal escape of macromolecules such as DNA are limited by their toxicity and by their ability to carry only limited ... More
Sec6/8 complexes on trans-Golgi network and plasma membrane regulate late stages of exocytosis in mammalian cells.
AuthorsYeaman C, Grindstaff KK, Wright JR, Nelson WJ
JournalJ Cell Biol
PubMed ID11696560
Sec6/8 complex regulates delivery of exocytic vesicles to plasma membrane docking sites, but how it is recruited to specific sites in the exocytic pathway is poorly understood. We identified an Sec6/8 complex on trans-Golgi network (TGN) and plasma membrane in normal rat kidney (NRK) cells that formed either fibroblast- (NRK-49F) ... More
Phosphoinositide 3-kinase regulates beta2-adrenergic receptor endocytosis by AP-2 recruitment to the receptor/beta-arrestin complex.
AuthorsNaga Prasad SV, Laporte SA, Chamberlain D, Caron MG, Barak L, Rockman HA
JournalJ Cell Biol
PubMed ID12163475
Internalization of beta-adrenergic receptors (betaARs) occurs by the sequential binding of beta-arrestin, the clathrin adaptor AP-2, and clathrin. D-3 phosphoinositides, generated by the action of phosphoinositide 3-kinase (PI3K) may regulate the endocytic process; however, the precise molecular mechanism is unknown. Here we demonstrate that betaARKinase1 directly interacts with the PIK ... More
Basic fibroblast growth factor stimulates surface expression and activity of Na(+)/H(+) exchanger NHE3 via mechanism involving phosphatidylinositol 3-kinase.
AuthorsJanecki AJ, Janecki M, Akhter S, Donowitz M
JournalJ Biol Chem
PubMed ID10713136
Na(+)/H(+) exchanger NHE3 is a plasma membrane (PM) protein, which contributes to Na(+) absorption in the intestine. Growth factors stimulate NHE3 via phosphatidylinositol 3-kinase (PI3-K), but mechanism of this process is not clear. To examine the hypothesis that growth factors stimulate NHE3 by modulating NHE3 recycling, and that PI3-K participates ... More
Endocytic recycling compartments altered in cisplatin-resistant cancer cells.
AuthorsLiang XJ, Mukherjee S, Shen DW, Maxfield FR, Gottesman MM
JournalCancer Res
PubMed ID16489040
The clinical utility of cisplatin to treat human malignancies is often limited by the development of drug resistance. We have previously shown that cisplatin-resistant human KB adenocarcinoma cells that are cross-resistant to methotrexate and heavy metals have altered endocytic recycling. In this work, we tracked lipids in the endocytic recycling ... More
The mouse SKD1, a homologue of yeast Vps4p, is required for normal endosomal trafficking and morphology in mammalian cells.
AuthorsYoshimori T, Yamagata F, Yamamoto A, Mizushima N, Kabeya Y, Nara A, Miwako I, Ohashi M, Ohsumi M, Ohsumi Y
JournalMol Biol Cell
PubMed ID10679028
The mouse SKD1 is an AAA-type ATPase homologous to the yeast Vps4p implicated in transport from endosomes to the vacuole. To elucidate a possible role of SKD1 in mammalian endocytosis, we generated a mutant SKD1, harboring a mutation (E235Q) that is equivalent to the dominant negative mutation (E233Q) in Vps4p. ... More
The p21 Rho-activating toxin cytotoxic necrotizing factor 1 is endocytosed by a clathrin-independent mechanism and enters the cytosol by an acidic-dependent membrane translocation step.
AuthorsContamin S, Galmiche A, Doye A, Flatau G, Benmerah A, Boquet P
JournalMol Biol Cell
PubMed ID10793151
Cytotoxic necrotizing factor 1 (CNF1), a protein produced by pathogenic strains of Escherichia coli, activates the p21 Rho-GTP-binding protein, inducing a profound reorganization of the actin cytoskeleton. CNF1 binds to its cell surface receptor on HEp-2 cells with high affinity (K(d) = 20 pM). In HEp-2 cells the action of ... More
Rab11BP/Rabphilin-11, a downstream target of rab11 small G protein implicated in vesicle recycling.
AuthorsMammoto A, Ohtsuka T, Hotta I, Sasaki T, Takai Y
JournalJ Biol Chem
PubMed ID10464283
Rab11 small G protein has been implicated in vesicle recycling, but its upstream regulators or downstream targets have not yet been identified. We isolated here a downstream target of Rab11, named rabphilin-11, from bovine brain. Moreover, we isolated from a rat brain cDNA library its cDNA, which encoded a protein ... More
Endocytosis resumes during late mitosis and is required for cytokinesis.
AuthorsSchweitzer JK, Burke EE, Goodson HV, D'Souza-Schorey C
JournalJ Biol Chem
PubMed ID16207714
Recent work has underscored the importance of membrane trafficking events during cytokinesis. For example, targeted membrane secretion occurs at the cleavage furrow in animal cells, and proteins that regulate endocytosis also influence the process of cytokinesis. Nonetheless, the prevailing dogma is that endosomal membrane trafficking ceases during mitosis and resumes ... More
Ligand-induced trafficking of the sphingosine-1-phosphate receptor EDG-1.
AuthorsLiu CH, Thangada S, Lee MJ, Van Brocklyn JR, Spiegel S, Hla T
JournalMol Biol Cell
PubMed ID10198065
The endothelial-derived G-protein-coupled receptor EDG-1 is a high-affinity receptor for the bioactive lipid mediator sphingosine-1-phosphate (SPP). In the present study, we constructed the EDG-1-green fluorescent protein (GFP) chimera to examine the dynamics and subcellular localization of SPP-EDG-1 interaction. SPP binds to EDG-1-GFP and transduces intracellular signals in a manner indistinguishable ... More
Endocytosis in skeletal muscle fibers.
AuthorsKaisto T, Rahkila P, Marjomäki V, Parton RG, Metsikkö K
JournalExp Cell Res
PubMed ID10585279
Defining the organization of endocytic pathway in multinucleated skeletal myofibers is crucial to understand the routing of membrane proteins, such as receptors and glucose transporters, through this system. Here we analyzed the organization of the endocytic trafficking pathways in isolated rat myofibers. We found that sarcolemmal-coated pits and transferrin receptors ... More
Mixed lineage kinase 2 interacts with clathrin and influences clathrin-coated vesicle trafficking.
AuthorsAkbarzadeh S, Ji H, Frecklington D, Marmy-Conus N, Mok YF, Bowes L, Devereux L, Linsenmeyer M, Simpson RJ, Dorow DS
JournalJ Biol Chem
PubMed ID12105200
Mixed lineage kinase 2 (MLK2) is a protein kinase that signals in the stress-activated Jun N-terminal kinase signal transduction pathway. We used immunoprecipitation and mass spectrometric analysis to identify MLK2-binding proteins in cell lines with inducible expression of green fluorescent protein-tagged MLK2. Here we report the identification of clathrin as ... More
Overexpression of ROCK in human breast cancer cells: evidence that ROCK activity mediates intracellular membrane traffic of lysosomes.
AuthorsNishimura Y, Itoh K, Yoshioka K, Tokuda K, Himeno M
JournalPathol Oncol Res
PubMed ID12858212
Small GTPase Rho and its downstream effectors, ROCK family of Rho-associated serine-threonine kinases, are thought to participate in cell morphology, motility, and tumor progression through regulating the rearrangement of actin cytoskeleton. Here we present evidence that transfection of human breast cancer cells with cDNA encoding a dominant active mutant of ... More
AP-2/Eps15 interaction is required for receptor-mediated endocytosis.
AuthorsBenmerah A, Lamaze C, Bègue B, Schmid SL, Dautry-Varsat A, Cerf-Bensussan N
JournalJ Cell Biol
PubMed ID9490719
We have previously shown that the protein Eps15 is constitutively associated with the plasma membrane adaptor complex, AP-2, suggesting its possible role in endocytosis. To explore the role of Eps15 and the function of AP-2/Eps15 association in endocytosis, the Eps15 binding domain for AP-2 was precisely delineated. The entire COOH-terminal ... More
The targeting of cystinosin to the lysosomal membrane requires a tyrosine-based signal and a novel sorting motif.
AuthorsCherqui S, Kalatzis V, Trugnan G, Antignac C
JournalJ Biol Chem
PubMed ID11150305
Cystinosis is a lysosomal transport disorder characterized by an accumulation of intra-lysosomal cystine. Biochemical studies showed that the lysosomal cystine transporter was distinct from the plasma membrane cystine transporters and that it exclusively transported cystine. The gene underlying cystinosis, CTNS, encodes a predicted seven-transmembrane domain protein called cystinosin, which is ... More
Actin-dependent propulsion of endosomes and lysosomes by recruitment of N-WASP.
AuthorsTaunton J, Rowning BA, Coughlin ML, Wu M, Moon RT, Mitchison TJ, Larabell CA
JournalJ Cell Biol
PubMed ID10662777
We examined the spatial and temporal control of actin assembly in living Xenopus eggs. Within minutes of egg activation, dynamic actin-rich comet tails appeared on a subset of cytoplasmic vesicles that were enriched in protein kinase C (PKC), causing the vesicles to move through the cytoplasm. Actin comet tail formation ... More
Differential localization of Rho GTPases in live cells: regulation by hypervariable regions and RhoGDI binding.
Determinants of membrane targeting of Rho proteins were investigated in live cells with green fluorescent fusion proteins expressed with or without Rho-guanine nucleotide dissociation inhibitor (GDI)alpha. The hypervariable region determined to which membrane compartment each protein was targeted. Targeting was regulated by binding to RhoGDI alpha in the case of ... More
Performance comparison between the high-speed Yokogawa spinning disc confocal system and single-point scanning confocal systems.
AuthorsWang E, Babbey CM, Dunn KW
JournalJ Microsc
PubMed ID15857376
Fluorescence microscopy of the dynamics of living cells presents a special challenge to a microscope imaging system, simultaneously requiring both high spatial resolution and high temporal resolution, but with illumination levels low enough to prevent fluorophore damage and cytotoxicity. We have compared the high-speed Yokogawa CSU10 spinning disc confocal system ... More
Bafilomycin A1 treatment retards transferrin receptor recycling more than bulk membrane recycling.
AuthorsPresley JF, Mayor S, McGraw TE, Dunn KW, Maxfield FR
JournalJ Biol Chem
PubMed ID9153255
Treatment of Chinese hamster ovary cells with the vacuolar proton pump inhibitor bafilomycin A1 causes a 2-fold retardation in the rate of recycling of transfected human transferrin receptors back to the cell surface as measured using biochemical assays (Johnson, L. S. , Dunn, K. W., Pytowski, B., and McGraw, T. ... More
Real time visualization of agonist-mediated redistribution and internalization of a green fluorescent protein-tagged form of the thyrotropin-releasing hormone receptor.
AuthorsDrmota T, Gould GW, Milligan G
JournalJ Biol Chem
PubMed ID9727016
The long isoform of the rat thyrotropin-releasing hormone receptor (TRHR) was modified by the addition of a vesicular stomatitis virus (VSV) epitope tag and green fluorescent protein (GFP). VSV-TRHR-GFP bound TRH with affinity similar to that of the unmodified receptor and stimulated [3H]inositol phosphate production. A clone stably expressing VSV-TRHR-GFP ... More
Co-trafficking of HFE, a nonclassical major histocompatibility complex class I protein, with the transferrin receptor implies a role in intracellular iron regulation.
AuthorsGross CN, Irrinki A, Feder JN, Enns CA
JournalJ Biol Chem
PubMed ID9705350
The mechanism by which a novel major histocompatibility complex class I protein, HFE, regulates iron uptake into the body is not known. HFE is the product of the gene that is mutated in >80% of hereditary hemochromatosis patients. It was recently found to coprecipitate with the transferrin receptor (Feder, J. ... More
Endocytic trafficking of megalin/RAP complexes: dissociation of the complexes in late endosomes.
AuthorsCzekay RP, Orlando RA, Woodward L, Lundstrom M, Farquhar MG
JournalMol Biol Cell
PubMed ID9188102
Megalin (gp330) is a member of the low-density lipoprotein receptor gene family. Like other members of the family, it is an endocytic receptor that binds a number of specific ligands. Megalin also binds the receptor-associated protein (RAP) that serves as an exocytic traffic chaperone and inhibits ligand binding to the ... More
Sorting of membrane components from endosomes and subsequent recycling to the cell surface occurs by a bulk flow process.
AuthorsMayor S, Presley JF, Maxfield FR
JournalJ Cell Biol
PubMed ID8509447
A central question in the endocytic process concerns the mechanism for sorting of recycling components (such as transferrin or low density lipoprotein receptors) from lysosomally directed components; membrane-associated molecules including receptors are generally directed towards the recycling pathway while the luminal content of sorting endosomes, consisting of the acid-released ligands, ... More
AuthorsBarlic J, Khandaker MH, Mahon E, Andrews J, DeVries ME, Mitchell GB, Rahimpour R, Tan CM, Ferguson SS, Kelvin DJ
JournalJ Biol Chem
PubMed ID10347185
The functional role of neutrophils during acute inflammatory responses is regulated by two high affinity interleukin-8 receptors (CXCR1 and CXCR2) that are rapidly desensitized and internalized upon binding their cognate chemokine ligands. The efficient re-expression of CXCR1 on the surface of neutrophils following agonist-induced internalization suggests that CXCR1 surface receptor ... More
Bacterial lipopolysaccharide can enter monocytes via two CD14-dependent pathways.
AuthorsKitchens RL, Wang P, Munford RS
JournalJ Immunol
PubMed ID9820530
Host recognition and disposal of LPS, an important Gram-negative bacterial signal molecule, may involve intracellular processes. We have therefore analyzed the initial pathways by which LPS, a natural ligand of glycosylphosphatidylinositol (GPI)-anchored CD14 (CD14-GPI), enters CD14-expressing THP-1 cells and normal human monocytes. Exposure of the cells to hypertonic medium obliterated ... More
Microtubule-dependent transport of secretory vesicles visualized in real time with a GFP-tagged secretory protein.
AuthorsWacker I, Kaether C, Krömer A, Migala A, Almers W, Gerdes HH
JournalJ Cell Sci
PubMed ID9224763
Biosynthetic transport from the trans-Golgi network (TGN) to the plasma membrane (PM) is mediated by secretory vesicles. We analyzed secretory vesicle transport in real time using a GFP-tagged secretory protein, hCgB-GFP, consisting of human chromogranin B (hCgB) and green fluorescent protein (GFP). The fusion protein was expressed transiently in Vero ... More
Clathrin and HA2 adaptors: effects of potassium depletion, hypertonic medium, and cytosol acidification.
AuthorsHansen SH, Sandvig K, van Deurs B
JournalJ Cell Biol
PubMed ID8458873
The effects of methods known to perturb endocytosis from clathrin-coated pits on the localization of clathrin and HA2 adaptors in HEp-2 carcinoma cells have been studied by immunofluorescence and ultrastructural immunogold microscopy, using internalization of transferrin as a functional assay. Potassium depletion, as well as incubation in hypertonic medium, remove ... More
Role of endocytosis in the internalization of spermidine-C(2)-BODIPY, a highly fluorescent probe of polyamine transport.
AuthorsSoulet D, Covassin L, Kaouass M, Charest-Gaudreault R, Audette M, Poulin R
JournalBiochem J
PubMed ID12097141
The mechanism of transmembrane polyamine internalization in mammalian cells remains unknown. A novel fluorescent spermidine conjugate [Spd-C(2)-BODIPY; N-(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-propionyl)-N'-(S -[spermidine-(N(4)-ethyl)]thioacetyl)ethylenediamine] was synthesized from N(4)-(mercaptoethyl)spermidine by a simple, one-step coupling procedure. In Chinese-hamster ovary (CHO) cells, Spd-C(2)-BODIPY accumulation was inhibited by exogenous putrescine, spermidine and spermine, was subject to feedback transport inhibition ... More
Multivesicular bodies in HEp-2 cells are maturing endosomes.
Conventional fluorescence microscopy of fixed HEp-2 cells as well as video microscopy of living cells incubated with transferrin-Texas Red (Tf-TxR) for < 60 min revealed distinct punctuate endosomal structures. Quantitative ultrastructural analysis using horseradish peroxidase (HRP) and cationized gold as tracers showed that spherical multivesicular bodies (MVBs) were the predominant ... More
Synaptic-like microvesicles of neuroendocrine cells originate from a novel compartment that is continuous with the plasma membrane and devoid of transferrin receptor.
AuthorsSchmidt A, Hannah MJ, Huttner WB
JournalJ Cell Biol
PubMed ID9128254
We have characterized the compartment from which synaptic-like microvesicles (SLMVs), the neuroendocrine counterpart of neuronal synaptic vesicles, originate. For this purpose we have exploited the previous observation that newly synthesized synaptophysin, a membrane marker of synaptic vesicles and SLMVs, is delivered to the latter organelles via the plasma membrane and ... More
A double leucine within the GLUT4 glucose transporter COOH-terminal domain functions as an endocytosis signal.
AuthorsCorvera S, Chawla A, Chakrabarti R, Joly M, Buxton J, Czech MP
JournalJ Cell Biol
PubMed ID7519625
The unique COOH-terminal 30-amino acid region of the adipocyte/skeletal muscle glucose transporter (GLUT4) appears to be a major structural determinant of this protein's perinuclear localization, from where it is redistributed to the cell surface in response to insulin. To test whether an underlying mechanism of this domain's function involves glucose ... More
Accumulation of cholera toxin and GM1 ganglioside in the early endosome of Niemann-Pick C1-deficient cells.
AuthorsSugimoto Y, Ninomiya H, Ohsaki Y, Higaki K, Davies JP, Ioannou YA, Ohno K
JournalProc Natl Acad Sci U S A
PubMed ID11675488
We investigated intracellular trafficking of GM1 ganglioside in Niemann-Pick C1 (NPC1)-deficient Chinese hamster ovary cells [NPC1(-) cells] by using cholera toxin (CT) as a probe. Both the holotoxin and the B subunit (CTB) accumulated in GM1-enriched intracellular vesicles of NPC1(-) cells. CTB-labeled vesicles contained the early endosome marker Rab5 but ... More
Differential roles of syntaxin 7 and syntaxin 8 in endosomal trafficking.
AuthorsPrekeris R, Yang B, Oorschot V, Klumperman J, Scheller RH
JournalMol Biol Cell
PubMed ID10564279
To understand molecular mechanisms that regulate the intricate and dynamic organization of the endosomal compartment, it is important to establish the morphology, molecular composition, and functions of the different organelles involved in endosomal trafficking. Syntaxins and vesicle-associated membrane protein (VAMP) families, also known as soluble N-ethylmaleimide-sensitive factor (NSF) attachment protein ... More
Potential sites of PI-3 kinase function in the endocytic pathway revealed by the PI-3 kinase inhibitor, wortmannin.
AuthorsShpetner H, Joly M, Hartley D, Corvera S
JournalJ Cell Biol
PubMed ID8647891
Previously we have shown that PDGF receptor mutants that do not bind PI-3 kinase internalize after ligand binding, but fail to downregulate and degrade. To define further the role of PI-3 kinase in trafficking processes in mammalian cells, we have investigated the effects of a potent inhibitor of PI-3 kinase ... More
Trafficking of Glut4-green fluorescent protein chimaeras in 3T3-L1 adipocytes suggests distinct internalization mechanisms regulating cell surface glut4 levels.
Insulin stimulates glucose transport in adipose and muscle tissue by stimulating the movement ('translocation') of an intracellular pool of glucose transporters (the Glut4 isoform) to the plasma membrane. We have engineered a series of chimaeras between Glut4 and green fluorescent protein (GFP) from Aequoria victoria and expressed these proteins in ... More
Inhibition of clathrin-dependent endocytosis has multiple effects on human rhinovirus serotype 2 cell entry.
AuthorsBayer N, Schober D, Hüttinger M, Blaas D, Fuchs R
JournalJ Biol Chem
PubMed ID11073943
Minor group human rhinoviruses (exemplified by human rhinovirus serotype 2 (HRV2)) use members of the low density lipoprotein receptor family for cell entry; all these receptors possess clathrin-coated pit localization signals. Viral infection should thus be inhibited under conditions of impaired clathrin-mediated endocytosis. However, Madshus et al. reported an increase ... More
Dynamic measurement of the pH of the Golgi complex in living cells using retrograde transport of the verotoxin receptor.
AuthorsKim JH, Lingwood CA, Williams DB, Furuya W, Manolson MF, Grinstein S
JournalJ Cell Biol
PubMed ID8830769
The B subunit of verotoxin (VT1B) from enterohemorrhagic Escherichia coli is responsible for the attachment of the holotoxin to the cell surface, by binding to the glycolipid, globotriaosyl ceramide. After receptor-mediated endocytosis, the toxin is targeted to the Golgi complex by a process of retrograde transport. We took advantage of ... More
Cellubrevin-targeted fluorescence uncovers heterogeneity in the recycling endosomes.
AuthorsTeter K, Chandy G, Quiñones B, Pereyra K, Machen T, Moore HP
JournalJ Biol Chem
PubMed ID9677389
The pH and trafficking of recycling endosomes have previously been studied using transferrin. We have used another approach, one in which the vesicle transport protein cellubrevin was appended with a luminal IgG epitope to allow targeting of fluorescein-5'-isothiocyanate (FITC)-labeled anti-IgG F(ab) antibodies to the recycling endosomes in living cells. FITC-F(ab) ... More
N-terminal tyrosine modulation of the endocytic adaptor function of the beta-arrestins.
AuthorsMarion S, Fralish GB, Laporte S, Caron MG, Barak LS
JournalJ Biol Chem
PubMed ID17456469
The highly homologous beta-arrestin1 and -2 adaptor proteins play important roles in the function of G protein-coupled receptors. Either beta-arrestin variant can function as a molecular chaperone for clathrin-mediated receptor internalization. This role depends primarily upon two distinct, contiguous C-terminal beta-arrestin motifs recognizing clathrin and the beta-adaptin subunit of AP2. ... More
Cationic albumin-conjugated pegylated nanoparticles allow gene delivery into brain tumors via intravenous administration.
AuthorsLu W, Sun Q, Wan J, She Z, Jiang XG
JournalCancer Res
PubMed ID17178885
Patients with malignant gliomas have a poor prognosis because these tumors do not respond well to conventional treatments. Studies of glioma xenografts suggest that they may be amenable to gene therapy with cytotoxic genes, such as the proapoptotic Apo2 ligand/tumor necrosis factor-related apoptosis-inducing ligand (Apo2L/TRAIL). Gene therapy of gliomas ideally ... More
Trafficking of cell surface beta-amyloid precursor protein: retrograde and transcytotic transport in cultured neurons.
AuthorsYamazaki T, Selkoe DJ, Koo EH
JournalJ Cell Biol
PubMed ID7721945
Amyloid beta-protein (A beta), the principal constituent of senile plaques seen in Alzheimer's disease (AD), is derived by proteolysis from the beta-amyloid precursor protein (beta PP). The mechanism of A beta production in neurons, which are hypothesized to be a rich source of A beta in brain, remains to be ... More
Differential localization of vesicular acetylcholine and monoamine transporters in PC12 cells but not CHO cells.
AuthorsLiu Y, Edwards RH
JournalJ Cell Biol
PubMed ID9362509
Previous studies have indicated that neuro-endocrine cells store monoamines and acetylcholine (ACh) in different secretory vesicles, suggesting that the transport proteins responsible for packaging these neurotransmitters sort to distinct vesicular compartments. Molecular cloning has recently demonstrated that the vesicular transporters for monoamines and ACh show strong sequence similarity, and studies ... More
Localization and insulin-regulated relocation of phosphoinositide 5-kinase PIKfyve in 3T3-L1 adipocytes.
AuthorsShisheva A, Rusin B, Ikonomov OC, DeMarco C, Sbrissa D
JournalJ Biol Chem
PubMed ID11112776
The mammalian phosphoinositide kinase PIKfyve catalyzes the synthesis of phosphatidylinositol 5-P and phosphatidylinositol 3,5-P(2), thought essential in cellular functions, including membrane trafficking. To discern the intracellular loci of PIKfyve products' formation, we have examined the localization of PIKfyve protein versus enzymatic activity and a possible acutely regulated redistribution in 3T3-L1 ... More
Transport of bacterial lipopolysaccharide to the golgi apparatus.
AuthorsThieblemont N, Wright SD
JournalJ Exp Med
PubMed ID10449523
Addition of lipopolysaccharide (LPS) to cells in the form of LPS-soluble (s)CD14 complexes induces strong cellular responses. During this process, LPS is delivered from sCD14 to the plasma membrane, and the cell-associated LPS is then rapidly transported to an intracellular site. This transport appears to be important for certain cellular ... More
Cellular and molecular barriers to gene transfer by a cationic lipid.
AuthorsZabner J, Fasbender AJ, Moninger T, Poellinger KA, Welsh MJ
JournalJ Biol Chem
PubMed ID7642560
Cationic lipids are widely used for gene transfer in vitro and show promise as a vector for in vivo gene therapy applications. However, there is limited understanding of the cellular and molecular mechanisms involved. We investigated the individual steps in cationic lipid-mediated gene transfer to cultured cell lines. We used ... More
TGF-β signaling is associated with endocytosis at the pocket region of the primary cilium.
Authors
JournalCell Rep
PubMed ID23746451
The F-BAR domain protein PACSIN2 associates with Rac1 and regulates cell spreading and migration.
Authors
JournalJ Cell Sci
PubMed ID21693584
Small molecule induced oligomerization, clustering and clathrin-independent endocytosis of the dopamine transporter.
Authors
JournalElife
PubMed ID29630493
Quantitative Profiling of the Lymph Node Clearance Capacity.
Authors
JournalSci Rep
PubMed ID30050160
Measles Virus Enters Breast and Colon Cancer Cell Lines through a PVRL4-Mediated Macropinocytosis Pathway.
Authors
JournalJ Virol
PubMed ID28250131
Loss of Bin1 Promotes the Propagation of Tau Pathology.
Authors
JournalCell Rep
PubMed ID27760323
Herpes Simplex Virus Entry by a Nonconventional Endocytic Pathway.
Authors
JournalJ Virol
PubMed ID33028710
A human polymorphism affects NEDD4L subcellular targeting by leading to two isoforms that contain or lack a C2 domain.
Authors
JournalBMC Cell Biol
PubMed ID19364400
Crossing the blood-brain-barrier with nanoligand drug carriers self-assembled from a phage display peptide.