Ubiquitination and proteasomal activity is required for transport of the EGF receptor to inner membranes of multivesicular bodies.
AuthorsLongva KE, Blystad FD, Stang E, Larsen AM, Johannessen LE, Madshus IH
JournalJ Cell Biol
PubMed ID11864992
'EGF, but not TGF alpha, efficiently induces degradation of the EGF receptor (EGFR). We show that EGFR was initially polyubiquitinated to the same extent upon incubation with EGF and TGF alpha, whereas the ubiquitination was more sustained by incubation with EGF than with TGF alpha. Consistently, the ubiquitin ligase c-Cbl ... More
Direct pathway from early/recycling endosomes to the Golgi apparatus revealed through the study of shiga toxin B-fragment transport.
AuthorsMallard F, Antony C, Tenza D, Salamero J, Goud B, Johannes L
JournalJ Cell Biol
PubMed ID9817755
'Shiga toxin and other toxins of this family can escape the endocytic pathway and reach the Golgi apparatus. To synchronize endosome to Golgi transport, Shiga toxin B-fragment was internalized into HeLa cells at low temperatures. Under these conditions, the protein partitioned away from markers destined for the late endocytic pathway ... More
Stonin 2: an adaptor-like protein that interacts with components of the endocytic machinery.
'Endocytosis of cell surface proteins is mediated by a complex molecular machinery that assembles on the inner surface of the plasma membrane. Here, we report the identification of two ubiquitously expressed human proteins, stonin 1 and stonin 2, related to components of the endocytic machinery. The human stonins are homologous ... More
A novel 14-kilodalton protein interacts with the mitogen-activated protein kinase scaffold mp1 on a late endosomal/lysosomal compartment.
AuthorsWunderlich W, Fialka I, Teis D, Alpi A, Pfeifer A, Parton RG, Lottspeich F, Huber LA
JournalJ Cell Biol
PubMed ID11266467
'We have identified a novel, highly conserved protein of 14 kD copurifying with late endosomes/lysosomes on density gradients. The protein, now termed p14, is peripherally associated with the cytoplasmic face of late endosomes/lysosomes in a variety of different cell types.In a two-hybrid screen with p14 as a bait, we identified ... More
Cytochalasins and colchicine increase the lateral mobility of human chorionic gonadotropin-occupied luteinizing hormone receptors on ovine luteal cells.
AuthorsRoess DA, Niswender GD, Barisas BG
JournalEndocrinology
PubMed ID3257187
'To determine whether cytoskeletal proteins restrict the mobility of LH-receptor complexes occupied by hCG, we measured the rate of lateral diffusion and the fraction of mobile receptor-hormone complexes by fluorescence photobleaching recovery on luteal cells treated with microtubule and microfilament disruptors and on portions of the plasma membrane separated from ... More
Abl tyrosine kinase regulates endocytosis of the epidermal growth factor receptor.
AuthorsTanos B, Pendergast AM
JournalJ Biol Chem
PubMed ID16943190
'Signal attenuation from ligand-activated epidermal growth factor receptor (EGFR) is mediated in part by receptor endocytosis and trafficking to the lysosomal degradative compartment. Uncoupling the activated EGFR from endocytosis and degradation has emerged as a mechanism for oncogenic activation of the EGFR. The Abl nonreceptor tyrosine kinase is activated by ... More
FCS cell surface measurements--photophysical limitations and consequences on molecular ensembles with heterogenic mobilities.
AuthorsWidengren J, Thyberg P
JournalCytometry A
PubMed ID16237686
'BACKGROUND: Fluorescence Correlation Spectroscopy is a powerful method to analyze densities and diffusive behavior of molecules in membranes, but effects of photodegradation can easily be overlooked. METHOD: Based on experimental photophysical parameters, calculations were performed to analyze the consequences of photobleaching in fluorescence correlation spectroscopy (FCS) cell surface experiments, covering ... More
The endocytosis of epidermal growth factor in A431 cells: a pH of microenvironment and the dynamics of receptor complex dissociation.
AuthorsSorkin AD, Teslenko LV, Nikolsky NN
JournalExp Cell Res
PubMed ID2894318
'The endocytosis and intracellular fate of epidermal growth factor (EGF) were studied in A431 cells. After 15-20 min of internalization at 37 degrees C, rhodamine-labeled EGF (EGF-Rh) accumulated into large juxtanuclear compartment consisting of closely related vesicles. This structure was shown to be localized in the para-Golgi region. Fluorescein-labeled transferrin ... More
Double-sided ubiquitin binding of Hrs-UIM in endosomal protein sorting.
AuthorsHirano S, Kawasaki M, Ura H, Kato R, Raiborg C, Stenmark H, Wakatsuki S
JournalNat Struct Mol Biol
PubMed ID16462748
'Hrs has an essential role in sorting of monoubiquitinated receptors to multivesicular bodies for lysosomal degradation, through recognition of ubiquitinated receptors by its ubiquitin-interacting motif (UIM). Here, we present the structure of a complex of Hrs-UIM and ubiquitin at 1.7-A resolution. Hrs-UIM forms a single alpha-helix, which binds two ubiquitin ... More
Systems biological analysis of epidermal growth factor receptor internalization dynamics for altered receptor levels.
AuthorsSchmidt-Glenewinkel H, Reinz E, Eils R, Brady NR,
JournalJ Biol Chem
PubMed ID19297331
'Epidermal growth factor (EGF) receptor (EGFR) overexpression is a hallmark of many cancers. EGFR endocytosis is a critical step in signal attenuation, raising the question of how receptor expression levels affect the internalization process. Here we combined quantitative experimental and mathematical modeling approaches to investigate the role of the EGFR ... More
Single-molecule analysis of epidermal growth factor signaling that leads to ultrasensitive calcium response.
AuthorsUyemura T, Takagi H, Yanagida T, Sako Y
JournalBiophys J
PubMed ID15749770
'Quantitative relationships between inputs and outputs of signaling systems are fundamental information for the understanding of the mechanism of signal transduction. Here we report the correlation between the number of epidermal growth factor (EGF) bindings and the response probability of intracellular calcium elevation. Binding of EGF molecules and changes of ... More
Epidermal growth factor receptor distribution during chemotactic responses.
AuthorsBailly M, Wyckoff J, Bouzahzah B, Hammerman R, Sylvestre V, Cammer M, Pestell R, Segall JE
JournalMol Biol Cell
PubMed ID11071913
'To determine the distribution of the epidermal growth factor (EGF) receptor (EGFR) on the surface of cells responding to EGF as a chemoattractant, an EGFR-green fluorescent protein chimera was expressed in the MTLn3 mammary carcinoma cell line. The chimera was functional and easily visualized on the cell surface. In contrast ... More
CIN85 phosphorylation is essential for EGFR ubiquitination and sorting into multivesicular bodies.
AuthorsSchroeder B, Srivatsan S, Shaw A, Billadeau D, McNiven MA,
JournalMol Biol Cell
PubMed ID22833562
Ubiquitination of the epidermal growth factor receptor (EGFR) by cbl and its cognate adaptor cbl-interacting protein of 85 kDa (CIN85) is known to play an essential role in directing this receptor to the lysosome for degradation. The mechanisms by which this ubiquitin modification is regulated are not fully defined, nor ... More
A novel endocytic mechanism of epidermal growth factor receptor sequestration and internalization.
Cells form transient, circular dorsal ruffles or "waves" in response to stimulation of receptor tyrosine kinases, including epidermal growth factor receptor (EGFR) or platelet-derived growth factor receptor. These dynamic structures progress inward on the dorsal surface and disappear, occurring concomitantly with a marked reorganization of F-actin. The cellular function of ... More
Reggies/flotillins regulate E-cadherin-mediated cell contact formation by affecting EGFR trafficking.
AuthorsSolis GP, Schrock Y, Hülsbusch N, Wiechers M, Plattner H, Stuermer CA,
JournalMol Biol Cell
PubMed ID22438585
The reggie/flotillin proteins are implicated in membrane trafficking and, together with the cellular prion protein (PrP), in the recruitment of E-cadherin to cell contact sites. Here, we demonstrate that reggies, as well as PrP down-regulation, in epithelial A431 cells cause overlapping processes and abnormal formation of adherens junctions (AJs). This ... More
Mitochondrially localized EGFR is independent of its endocytosis and associates with cell viability.
AuthorsYao Y, Wang G, Li Z, Yan B, Guo Y, Jiang X, Xi Z,
JournalActa Biochim Biophys Sin (Shanghai)
PubMed ID20929928
The molecular mechanism underlying epidermal growth factor receptor (EGFR) localization in mitochondria remains largely unknown. Using immune electron microscopy, we validated that EGFR could be localized on either the outer or the inner membrane of mitochondria. Mutant receptor lacked amino acids 646-660 was flawed in migration onto the organelles, whereas ... More
Decorin evokes protracted internalization and degradation of the epidermal growth factor receptor via caveolar endocytosis.
Decorin inhibits the epidermal growth factor receptor (EGFR) by down-regulating its tyrosine kinase activity, thereby blocking the growth of a variety of transformed cells and tumor xenografts. In this study we provide evidence that decorin directly binds to the EGFR causing its dimerization, internalization, and ultimately its degradation. Using various ... More
Large deletions in the cytoplasmic kinase domain of the epidermal growth factor receptor do not affect its laternal mobility.
AuthorsLivneh E, Benveniste M, Prywes R, Felder S, Kam Z, Schlessinger J
JournalJ Cell Biol
PubMed ID3015981
The lateral diffusion coefficients of various epidermal growth factor (EGF) receptor mutants with increasing deletions in their carboxy-terminal cytoplasmic domain were compared. A full size cDNA construct of human EGF receptor and different deletion constructs were expressed in monkey COS cells. The EGF receptor mutants expressed on the cell surface ... More
High affinity epidermal growth factor receptors on the surface of A431 cells have restricted lateral diffusion.
AuthorsRees AR, Gregoriou M, Johnson P, Garland PB
JournalEMBO J
PubMed ID6090128
Rhodamine-labelled epidermal growth factor (Rh-EGF) was shown to bind to A431 cells grown at low density both to a small number of high affinity receptors (KD = 2.8 X 10(-10) M; fraction of total binding sites approximately 0.12) and also to a large number of low affinity receptors (KD = ... More
Overexpression of epidermal growth factor receptor in NIH-3T3-transfected cells slows its lateral diffusion and rate of endocytosis.
AuthorsBenveniste M, Livneh E, Schlessinger J, Kam Z
JournalJ Cell Biol
PubMed ID3133376
Interactions between membrane proteins are believed to be important for the induction of transmembrane signaling. Endocytosis is one of the responses which is regulated by both intracellular and extracellular signals. To study such interactions, we have measured the lateral mobility and rate of endocytosis of epidermal growth factor receptor in ... More
Oligomerization of epidermal growth factor receptors on A431 cells studied by time-resolved fluorescence imaging microscopy. A stereochemical model for tyrosine kinase receptor activation.
AuthorsGadella TW, Jovin TM
JournalJ Cell Biol
PubMed ID7790353
The aggregation states of the epidermal growth factor receptor (EGFR) on single A431 human epidermoid carcinoma cells were assessed with two new techniques for determining fluorescence resonance energy transfer: donor photobleaching fluorescence resonance energy transfer (pbFRET) microscopy and fluorescence lifetime imaging microscopy (FLIM). Fluorescein-(donor) and rhodamine-(acceptor) labeled EGF were bound ... More
SH2-containing 5'-inositol phosphatase, SHIP2, regulates cytoskeleton organization and ligand-dependent down-regulation of the epidermal growth factor receptor.
AuthorsPrasad NK, Decker SJ
JournalJ Biol Chem
PubMed ID15668240
Phosphoinositide lipid second messengers are integral components of signaling pathways mediated by insulin, growth factors, and integrins. SHIP2 dephosphorylates phosphatidylinositol 3,4,5-trisphosphate generated by the activated phosphatidylinositol 3'-kinase. SHIP2 down-regulates insulin signaling and is present at higher levels in diabetes and obesity. SHIP2 associates with p130Cas and filamin, regulators of cell ... More
Characterization of internalization and endosome formation of epidermal growth factor in transfected NIH-3T3 cells by computerized image-intensified three-dimensional fluorescence microscopy.
AuthorsBenveniste M, Schlessinger J, Kam Z
JournalJ Cell Biol
PubMed ID2808521
Computerized image-intensified fluorescence microscopy has been used to quantify routing and subcellular concentrations of rhodaminated EGF (Rh-EGF) during its receptor-mediated endocytosis in two transfected NIH-3T3 cell lines expressing 2 X 10(5) and 1.5 X 10(6) receptors per cell, respectively. A series of images were digitized by focusing at different depths ... More
Rab7: a key to lysosome biogenesis.
AuthorsBucci C, Thomsen P, Nicoziani P, McCarthy J, van Deurs B
JournalMol Biol Cell
PubMed ID10679007
The molecular machinery behind lysosome biogenesis and the maintenance of the perinuclear aggregate of late endocytic structures is not well understood. A likely candidate for being part of this machinery is the small GTPase Rab7, but it is unclear whether this protein is associated with lysosomes or plays any role ... More
Generation of endogenous pH-sensitive EGF receptor and its application in high-throughput screening for proteins involved in clathrin-mediated endocytosis.
Authors
JournalElife
PubMed ID31066673
UVRAG mutations associated with microsatellite unstable colon cancer do not affect autophagy.
Authors
JournalAutophagy
PubMed ID20724836
Inhibition of Cyclin-Dependent Kinase 5: A Strategy to Improve Sorafenib Response in Hepatocellular Carcinoma Therapy.
Authors
JournalHepatology
PubMed ID30033593
Comprehensive Model for Epidermal Growth Factor Receptor Ligand Binding Involving Conformational States of the Extracellular and the Kinase Domains.
Authors
JournalFront Cell Dev Biol
PubMed ID32850868
Analytical characterization of plasma membrane-derived vesicles produced via osmotic and chemical vesiculation.
Authors
JournalBiochim Biophys Acta
PubMed ID25896659
Alteration of the late endocytic pathway in Charcot-Marie-Tooth type 2B disease.
Authors
JournalCell Mol Life Sci
PubMed ID32280996
Tannins from Syzygium guineense suppress Wnt signaling and proliferation of Wnt-dependent tumors through a direct effect on secreted Wnts.