Nocodazole-dependent transport, and brefeldin A--sensitive processing and sorting, of newly synthesized membrane proteins in cultured neurons.
AuthorsCid-Arregui A, Parton RG, Simons K, Dotti CG
JournalJ Neurosci
PubMed ID7790910
The envelope glycoproteins of Semliki Forest virus (SFV), Vesicular Stomatitis virus (VSV), and Influenza Fowl Plague virus (FPV) are vectorially targeted in neurons to the plasma membrane of dendrites (SFV and VSV) and axons (FPV). To gain insight into the mechanisms responsible for such polarized delivery we have examined the ... More
Mammalian Cdc42 is a brefeldin A-sensitive component of the Golgi apparatus.
AuthorsErickson JW, Zhang C, Kahn RA, Evans T, Cerione RA
JournalJ Biol Chem
PubMed ID8900167
'In this study, we have used immunocytochemical and fractionation approaches to provide a description of the localization of the mammalian Cdc42 protein (designated Cdc42Hs) in vivo. A specific anti-peptide antibody was generated against the C-terminal region of Cdc42Hs. Using affinity-purified preparations of this antibody in indirect immunofluorescence experiments, Cdc42Hs was ... More
Molecular cloning of a novel 97-kd Golgi complex autoantigen associated with Sjögren's syndrome.
'OBJECTIVE: To identify a Golgi complex autoantigen bound by Sjögren''s syndrome (SS) autoantibodies. METHODS: Serum from a patient with secondary SS and anti-Golgi antibodies was used as a probe to isolate a complementary DNA (cDNA) insert from a HeLa cDNA library. RESULTS: A 3.7-kb cDNA encoding a 56-kd recombinant protein ... More
An ordered inheritance strategy for the Golgi apparatus: visualization of mitotic disassembly reveals a role for the mitotic spindle.
AuthorsShima DT, Cabrera-Poch N, Pepperkok R, Warren G
JournalJ Cell Biol
PubMed ID9585414
'During mitosis, the ribbon of the Golgi apparatus is transformed into dispersed tubulo-vesicular membranes, proposed to facilitate stochastic inheritance of this low copy number organelle at cytokinesis. Here, we have analyzed the mitotic disassembly of the Golgi apparatus in living cells and provide evidence that inheritance is accomplished through an ... 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
Kinesin-mediated organelle translocation revealed by specific cellular manipulations.
AuthorsFeiguin F, Ferreira A, Kosik KS, Caceres A
JournalJ Cell Biol
PubMed ID7962067
'The distribution of membrane-bound organelles was studied in cultured hippocampal neurons after antisense oligonucleotide suppression of the kinesin-heavy chain (KHC). We observed reduced 3,3''-dihexyloxacarbocyanine iodide (DiOC6(3)) fluorescent staining in neurites and growth cones. In astrocytes, KHC suppression results in the disappearance of the DiOC6(3)-positive reticular network from the cell periphery, ... More
ZnT7, a novel mammalian zinc transporter, accumulates zinc in the Golgi apparatus.
AuthorsKirschke CP, Huang L,
JournalJ Biol Chem
PubMed ID12446736
'ZnT7, a novel member of the zinc transporter (ZnT) family, was identified by searching the expressed sequence tag (EST) databases with the amino acid sequence of ZnT1. Like the other ZnT proteins, the protein (387 amino acids) predicted from this gene contains six transmembrane domains and a histidine-rich loop between ... More
The effect of Golgi depletion on exocytic transport.
AuthorsPelletier L, Jokitalo E, Warren G
JournalNat Cell Biol
PubMed ID11056540
'Here we evaluate the idea that the Golgi is in dynamic equilibrium with the endoplasmic reticulum (ER). In cytoplasts that lack the Golgi apparatus, no regrowth of the Golgi is observed, nor is any transport from the ER to the cell surface detected. However, introduction of the smallest measurable amount ... More
Forskolin inhibits and reverses the effects of brefeldin A on Golgi morphology by a cAMP-independent mechanism.
'Brefeldin A (BFA) causes rapid redistribution of Golgi proteins into the ER, leaving no definable Golgi apparatus, and blocks transport of proteins into post-Golgi compartments in the cell. In this study we follow the disassembly of the Golgi apparatus in BFA-treated, living cells labeled with NBD-ceramide and demonstrate that forskolin ... More
A furin-like convertase mediates propeptide cleavage of BACE, the Alzheimer's beta -secretase.
AuthorsBennett BD, Denis P, Haniu M, Teplow DB, Kahn S, Louis JC, Citron M, Vassar R
JournalJ Biol Chem
PubMed ID10956649
'The novel transmembrane aspartic protease BACE (for Beta-site APP Cleaving Enzyme) is the beta-secretase that cleaves amyloid precursor protein to initiate beta-amyloid formation. As such, BACE is a prime therapeutic target for the treatment of Alzheimer''s disease. BACE, like other aspartic proteases, has a propeptide domain that is removed to ... More
A pore-forming toxin interacts with a GPI-anchored protein and causes vacuolation of the endoplasmic reticulum.
AuthorsAbrami L, Fivaz M, Glauser PE, Parton RG, van der Goot FG
JournalJ Cell Biol
PubMed ID9456314
'In this paper, we have investigated the effects of the pore-forming toxin aerolysin, produced by Aeromonas hydrophila, on mammalian cells. Our data indicate that the protoxin binds to an 80-kD glycosyl-phosphatidylinositol (GPI)-anchored protein on BHK cells, and that the bound toxin is associated with specialized plasma membrane domains, described as ... More
ADP-Ribosylation factor 1 (ARF1) regulates recruitment of the AP-3 adaptor complex to membranes.
AuthorsOoi CE, Dell'Angelica EC, Bonifacino JS
JournalJ Cell Biol
PubMed ID9679139
'Small GTP-binding proteins such as ADP- ribosylation factor 1 (ARF1) and Sar1p regulate the membrane association of coat proteins involved in intracellular membrane trafficking. ARF1 controls the clathrin coat adaptor AP-1 and the nonclathrin coat COPI, whereas Sar1p controls the nonclathrin coat COPII. In this study, we demonstrate that membrane ... More
Stimulation of sphingomyelin biosynthesis by brefeldin A and sphingomyelin breakdown by okadaic acid treatment of rat hepatocytes.
AuthorsHatch GM, Vance DE
JournalJ Biol Chem
PubMed ID1618752
'Studies on sphingomyelin metabolism in rat hepatocytes were facilitated by the use of choline-deficient cells which allowed for the rapid labeling of phosphatidylcholine and as a result sphingomyelin. Pulse and pulse-chase studies with [methyl-3H]choline and [methyl-3H]methionine demonstrated that both compounds were effectively used for sphingomyelin biosynthesis and that newly made ... More
Differential effects of brefeldin A on transport of secretory and lysosomal proteins.
AuthorsStrous GJ, van Kerkhof P, van Meer G, Rijnboutt S, Stoorvogel W
JournalJ Biol Chem
PubMed ID8428908
'Brefeldin A (BFA) rapidly blocks anterograde exocytotic transport through the Golgi complex. Sustained retrograde traffic induced by brefeldin A causes redistribution of constituents of the Golgi, but not the trans-Golgi network (TGN), to the endoplasmic reticulum (ER). In the present study on HepG2 cells, we have observed a differential effect ... More
Visualization and characterization of respiratory syncytial virus F-specific CD8(+) T cells during experimental virus infection.
AuthorsChang J, Srikiatkhachorn A, Braciale TJ
JournalJ Immunol
PubMed ID11591747
'CTL play a major role in the clearance of respiratory syncytial virus (RSV) during experimental pulmonary infection. The fusion (F) glycoprotein of RSV is a protective Ag that elicits CTL and Ab response against RSV infection in BALB/c mice. We used the strategy of screening a panel of overlapping synthetic ... More
Golgi tubule traffic and the effects of brefeldin A visualized in living cells.
AuthorsSciaky N, Presley J, Smith C, Zaal KJ, Cole N, Moreira JE, Terasaki M, Siggia E, Lippincott-Schwartz J
JournalJ Cell Biol
PubMed ID9382862
'The Golgi complex is a dynamic organelle engaged in both secretory and retrograde membrane traffic. Here, we use green fluorescent protein-Golgi protein chimeras to study Golgi morphology in vivo. In untreated cells, membrane tubules were a ubiquitous, prominent feature of the Golgi complex, serving both to interconnect adjacent Golgi elements ... More
Microtubule-dependent retrograde transport of proteins into the ER in the presence of brefeldin A suggests an ER recycling pathway.
AuthorsLippincott-Schwartz J, Donaldson JG, Schweizer A, Berger EG, Hauri HP, Yuan LC, Klausner RD
JournalCell
PubMed ID2178778
'Characteristics of brefeldin A (BFA)-induced redistribution of Golgi proteins into the endoplasmic reticulum (ER) and its relationship to an ER retrieval pathway were investigated. Retrograde movement of Golgi proteins into the ER occurred via long, tubulovesicular processes extending out of the Golgi along microtubules. Microtubule-disrupting agents (i.e., nocodazole), energy poisons, ... More
Brefeldin A reversibly inhibits secretion in Saccharomyces cerevisiae.
AuthorsShah N, Klausner RD
JournalJ Biol Chem
PubMed ID8449896
'Brefeldin A has proven to be a useful pharmacologic tool, which, when added to mammalian cells, results in a block in secretion as well as the structural disruption of specific intracellular organelles. In spite of our understanding of some of the biochemistry underlying the action of brefeldin A, the most ... More
ADP-ribosylation factor 6 regulates a novel plasma membrane recycling pathway.
AuthorsRadhakrishna H, Donaldson JG
JournalJ Cell Biol
PubMed ID9314528
'ADP-ribosylation factor (ARF) 6 localizes to the plasma membrane (PM) in its GTP state and to a tubulovesicular compartment in its GDP state in HeLa cells that express wild-type or mutant forms of this GTPase. Aluminum fluoride (AlF) treatment of ARF6-transfected cells redistributes ARF6 to the PM and stimulates the ... More
Evidence for a cholesterol transport pathway from lysosomes to endoplasmic reticulum that is independent of the plasma membrane.
AuthorsUnderwood KW, Jacobs NL, Howley A, Liscum L
JournalJ Biol Chem
PubMed ID9461625
'We have studied the movement of low density lipoprotein (LDL)-derived cholesterol in cultured Chinese hamster ovary cells. Our hypothesis is that when LDL cholesterol is effluxed from lysosomes, the bulk of LDL cholesterol is mobilized to the plasma membrane, while another pathway delivers LDL cholesterol from lysosomes to acyl-CoA/cholesterol acyltransferase ... More
Dynamics of transitional endoplasmic reticulum sites in vertebrate cells.
AuthorsHammond AT, Glick BS
JournalMol Biol Cell
PubMed ID10982397
'A typical vertebrate cell contains several hundred sites of transitional ER (tER). Presumably, tER sites generate elements of the ER-Golgi intermediate compartment (ERGIC), and ERGIC elements then generate Golgi cisternae. Therefore, characterizing the mechanisms that influence tER distribution may shed light on the dynamic behavior of the Golgi. We explored ... More
A novel class of clathrin-coated vesicles budding from endosomes.
AuthorsStoorvogel W, Oorschot V, Geuze HJ
JournalJ Cell Biol
PubMed ID8567724
'Clathrin-coated vesicles transport selective integral membrane proteins from the plasma membrane to endosomes and from the TGN to endosomes. Recycling of proteins from endosomes to the plasma membrane occurs via unidentified vesicles. To study this pathway, we used a novel technique that allows for the immunoelectron microscopic examination of transferrin ... More
Insulin-sensitive association of GLUT-4 with endocytic clathrin-coated vesicles revealed with the use of brefeldin A.
AuthorsChakrabarti R, Buxton J, Joly M, Corvera S
JournalJ Biol Chem
PubMed ID8132512
'The interaction of the adipocyte/skeletal muscle glucose transporter (GLUT-4) with clathrin lattices may be important in maintaining its intracellular distribution. To better understand the role of clathrin lattices in the sorting of GLUT-4, we have attempted to determine the cellular origin of clathrin-coated vesicles (CCVs) that contain this transporter. The ... More
Inhibition by brefeldin A of a Golgi membrane enzyme that catalyses exchange of guanine nucleotide bound to ARF.
AuthorsHelms JB, Rothman JE
JournalNature
PubMed ID1448152
'A wide variety of membrane transformations important in intracellular transport are inhibited by the fungal metabolite brefeldin A (refs 1-4), implying that the target for this drug is central to the formation and maintenance of subcellular compartments. Brefeldin A added to cells causes the rapid and reversible dissociation of a ... More
Brefeldin A does not inhibit the movement of phosphatidylethanolamine from its sites for synthesis to the cell surface.
AuthorsVance JE, Aasman EJ, Szarka R
JournalJ Biol Chem
PubMed ID2022641
'Brefeldin A, a fungal metabolite which interrupts trafficking of proteins via the Golgi by causing disassembly of the Golgi stacks, has been used to investigate the mechanism of movement of phosphatidylethanolamine (PtdEtn) from its sites of synthesis to the cell surface. PtdEtn is made in hepatocytes by two major pathways, ... More
Developmental induction of Golgi structure and function in the primitive eukaryote Giardia lamblia.
AuthorsLuján HD, Marotta A, Mowatt MR, Sciaky N, Lippincott-Schwartz J, Nash TE
JournalJ Biol Chem
PubMed ID7876232
'A fundamental characteristic of eukaryotic cells is the presence of membrane-bound compartments and membrane transport pathways in which the Golgi complex plays a central role in the selective processing, sorting, and secretion of proteins. The parasitic protozoan Giardia lamblia belongs to the earliest identified lineage among eukaryotes and therefore offers ... More
Effects of brefeldin A on endocytosis, transcytosis and transport to the Golgi complex in polarized MDCK cells.
AuthorsPrydz K, Hansen SH, Sandvig K, van Deurs B
JournalJ Cell Biol
PubMed ID1400572
'We have studied the effects of brefeldin A (BFA) on endocytosis and intracellular traffic in polarized MDCK cells by using the galactose-binding protein toxin ricin as a membrane marker and HRP as a marker of fluid phase transport. We found that BFA treatment rapidly increased apical endocytosis of both ricin ... More
ARF is required for maintenance of yeast Golgi and endosome structure and function.
AuthorsGaynor EC, Chen CY, Emr SD, Graham TR
JournalMol Biol Cell
PubMed ID9487133
'ADP ribosylation factor (ARF) is thought to play a critical role in recruiting coatomer (COPI) to Golgi membranes to drive transport vesicle budding. Yeast strains harboring mutant COPI proteins exhibit defects in retrograde Golgi to endoplasmic reticulum protein transport and striking cargo-selective defects in anterograde endoplasmic reticulum to Golgi protein ... More
Brefeldin A promotes hydrolysis of sphingomyelin.
AuthorsLinardic CM, Jayadev S, Hannun YA
JournalJ Biol Chem
PubMed ID1634530
'The hydrolysis of sphingomyelin (SM) is a key reaction in the "sphingomyelin cycle," which plays a role in the regulation of cell proliferation and differentiation (Okazaki, T., Bell, R. M., and Hannun, Y. A. (1989) J. Biol. Chem. 264, 19076-19080). SM is produced from endoplasmic reticulum-derived ceramide and is delivered ... More
The ubiquitin conjugation system is required for ligand-induced endocytosis and degradation of the growth hormone receptor.
AuthorsStrous GJ, van Kerkhof P, Govers R, Ciechanover A, Schwartz AL
JournalEMBO J
PubMed ID8670885
'The ubiquitin-dependent protein degradation system has recently been implicated in downregulation of signal transducing receptors. Growth hormone receptor (GHR) cDNA was transfected into Chinese hamster ovary cells, which exhibit a temperature-sensitive defect in ubiquitin conjugation (CHO-ts20), as well as into wild-type cells (CHO-E36). Upon binding of growth hormone (GH), two ... More
Effects of brefeldin A on the processing of viral envelope glycoproteins in murine erythroleukemia cells.
AuthorsUlmer JB, Palade GE
JournalJ Biol Chem
PubMed ID1851169
'This paper documents the effects of brefeldin A (BFA) on the processing and transport of viral envelope glycoproteins in a retrovirus-transformed murine erythroleukemia (MEL) cell line. BFA is a fungal metabolite that disrupts intracellular membrane traffic at the endoplasmic reticulum (ER)-Golgi complex junction. In MEL cells, BFA inhibited the processing ... More
Novel blockade by brefeldin A of intracellular transport of secretory proteins in cultured rat hepatocytes.
AuthorsMisumi Y, Misumi Y, Miki K, Takatsuki A, Tamura G, Ikehara Y
JournalJ Biol Chem
PubMed ID2426273
'We examined the effect of brefeldin A, an antiviral antibiotic, on protein synthesis, intracellular processing, and secretion in primary culture of rat hepatocytes. The secretion was strongly blocked by the drug at 1 microgram/ml and higher concentrations, while the protein synthesis was maintained fairly well. Pulse-chase experiments with [35S]methionine demonstrated ... More
Brefeldin A inhibits insulin-induced glucose transport stimulation and GLUT4 recruitment in rat adipocytes.
AuthorsLachaal M, Moronski C, Liu H, Jung CY
JournalJ Biol Chem
PubMed ID8089139
'GLUT4, the major insulin-responsive glucose transporter isoform in rat adipocytes, rapidly recycles between an intracellular pool and the plasma membrane in the basal and insulin-stimulated states. To gain insight into the route of this GLUT4 recycling, we studied the effects of brefeldin A (BFA) on glucose transport and glucose transporter ... More
DS28-6, a temperature-sensitive mutant of Chinese hamster ovary cells, expresses key phenotypic changes associated with brefeldin A treatment.
AuthorsZuber C, Roth J, Misteli T, Nakano A, Moremen K
JournalProc Natl Acad Sci U S A
PubMed ID1946407
'The temperature-sensitive Chinese hamster ovary (CHO) cell mutant DS28-6 has been previously shown to be pleiotropically defective in protein secretion. We have examined the mutant cells to determine the intracellular site of the block in secretion. By transmission electron microscopy a time-dependent disassembly of the Golgi apparatus was found under ... More
Golgi spectrin: identification of an erythroid beta-spectrin homolog associated with the Golgi complex.
AuthorsBeck KA, Buchanan JA, Malhotra V, Nelson WJ
JournalJ Cell Biol
PubMed ID7962054
'Spectrin is a major component of a membrane-associated cytoskeleton involved in the maintenance of membrane structural integrity and the generation of functionally distinct membrane protein domains. Here, we show that a homolog of erythrocyte beta-spectrin (beta I sigma*) co-localizes with markers of the Golgi complex in a variety of cell ... More
Brefeldin A enhances receptor-mediated transcytosis of transferrin in filter-grown Madin-Darby canine kidney cells.
AuthorsWan J, Taub ME, Shah D, Shen WC
JournalJ Biol Chem
PubMed ID1618847
'The effects of brefeldin A (BFA) on transferrin (Tf) transcellular transport, Tf receptor (TfR) distribution, and TfR-mediated endocytosis in filter-grown Madin-Darby canine kidney (MDCK) cells were studied. BFA (1.6 micrograms/ml) markedly enhanced the transcytosis of 125I-labeled Tf (125I-Tf) in both apical-to-basal and basal-to-apical directions; yet, BFA did not enhance the ... More
Ceramide reverses brefeldin A (BFA) resistance in BFA-resistant cell lines.
AuthorsOda T, Chen CH, Wu HC
JournalJ Biol Chem
PubMed ID7876158
'We have found that C6 ceramide, a cell-permeable ceramide analog, partially restored the brefeldin A (BFA) sensitivity in a BFA-resistant mutant of Vero cells (BER-40) and in the naturally BFA-resistant Madin-Darby canine kidney (MDCK) cells. Incubation of BER-40 and MDCK cells with low concentrations of C6 ceramide resulted in (i) ... More
Both alpha 1A- and alpha 2A-adrenoreceptor subtypes stimulate voltage-operated L-type calcium channels in rat portal vein myocytes. Evidence for two distinct transduction pathways.
AuthorsLeprêtre N, Mironneau J, Morel JL
JournalJ Biol Chem
PubMed ID7961939
'Previously, we have shown that myocytes from rat portal vein express alpha 1A-adrenoreceptors that couple with a Gq/G11-protein to stimulate phosphoinositide turnover and release of calcium from intracellular stores. The purpose of this study was to investigate the contribution of both alpha 1- and alpha 2-adrenoreceptor subtypes in inducing stimulation ... More
A recycling pathway between the endoplasmic reticulum and the Golgi apparatus for retention of unassembled MHC class I molecules.
'Assembly of class I major histocompatibility complex (MHC) molecules involves the interaction of two distinct polypeptides (the heavy and light chains) with peptide antigen. Cell lines synthesizing both chains but expressing low levels of MHC class I molecules on their surface as a result of a failure in assembly and ... More
VAMP2, but not VAMP3/cellubrevin, mediates insulin-dependent incorporation of GLUT4 into the plasma membrane of L6 myoblasts.
AuthorsRandhawa VK, Bilan PJ, Khayat ZA, Daneman N, Liu Z, Ramlal T, Volchuk A, Peng XR, Coppola T, Regazzi R, Trimble WS, Klip A
JournalMol Biol Cell
PubMed ID10888677
'Like neuronal synaptic vesicles, intracellular GLUT4-containing vesicles must dock and fuse with the plasma membrane, thereby facilitating insulin-regulated glucose uptake into muscle and fat cells. GLUT4 colocalizes in part with the vesicle SNAREs VAMP2 and VAMP3. In this study, we used a single-cell fluorescence-based assay to compare the functional involvement ... More
Guanosine nucleotides modulate the inhibitory effect of brefeldin A on protein secretion.
AuthorsZeuzem S, Zimmermann P, Schulz I
JournalFEBS Lett
PubMed ID1908795
'Brefeldin A (BFA) causes rapid redistribution of Golgi proteins into the endoplasmic reticulum (ER), leaving no definable Golgi-apparatus, and blocks transport of proteins from the ER to distal secretory compartments of the cell. Using pulse-chase experiments the present study shows that BFA (1 microgram/ml) inhibits basal and CCK-stimulated protein secretion ... More
Brefeldin A renders Chinese hamster ovary cells insensitive to transcriptional suppression by 25-hydroxycholesterol.
AuthorsRidgway ND, Lagace TA
JournalJ Biol Chem
PubMed ID7713903
'The effect of disruption of the Golgi apparatus on 25-hydroxycholesterol-mediated transcriptional suppression and activation of acyl-CoA:cholesterol acyltransferase was examined. In Chinese hamster ovary (CHO) cells, brefeldin A (BFA) caused dose-dependent inhibition of 25-hydroxycholesterol-mediated suppression of mRNAs for four sterol-regulated genes: 3-hydroxy-3-methylglutaryl (HMG)-CoA reductase, HMG-CoA synthase, farnesyl-diphosphate synthase, and the low ... More
The therapeutic effect of lipo PGE1 on diabetic neuropathy-changes in endothelin and various angiopathic factors.
AuthorsItoh Y, Yasui T, Kakizawa H, Makino M, Fujiwara K, Kato T, Imamura S, Yamamoto K, Hishida H, Nakai A, Itoh M, Nagasaka A
JournalProstaglandins Other Lipid Mediat
PubMed ID11577785
'A high blood concentration of endothelin (ET)-1 may participate in the onset and progress of diabetic microangiopathy, resulting in neuropathy. We examined the therapeutic effects of prostaglandin E1 (PGE1), which possesses both a peripheral vasodilating action and inhibition of platelet aggregation, on diabetic microangiopathy. Increases in both skin temperature and ... More
ATP-dependent translocation of ricin across the membrane of purified endosomes.
AuthorsBeaumelle B, Alami M, Hopkins CR
JournalJ Biol Chem
PubMed ID7901210
'Ricin translocation was demonstrated (using both fluorescence- and radiolabel-based assays) across the membrane of endosomes purified from mouse lymphocytes. Selectivity of the process was shown by the absence of translocation activity of transferrin and horseradish peroxidase used as membrane-bound and fluid-phase endosome labels, respectively. Endocytosed 125I-ricin translocation was found to ... More
Temporal down-regulation of Fc gamma RIII expression and Fc gamma receptor-mediated phagocytosis in human monocyte-derived macrophages induced by TNF-alpha and IL-1 beta.
AuthorsLiao G, Simon SR
JournalJ Leukoc Biol
PubMed ID8195695
'Effects of tumor necrosis factor (TNF-alpha) and interleukin-1 beta (IL-1 beta) on the expression of Fc gamma receptors (Fc gamma Rs) and on Fc gamma R-mediated phagocytosis in human monocyte-derived macrophages (MDMs) have been determined. Treatment of MDMs with TNF-alpha (400 pg/ml) or IL-1 beta (400 pg/ml) for 80 min ... More
The effects of brefeldin A on the glucose transport system in rat adipocytes. Implications regarding the intracellular locus of insulin-sensitive Glut4.
AuthorsBao S, Smith RM, Jarett L, Garvey WT
JournalJ Biol Chem
PubMed ID8530430
'Insulin activates glucose transport by recruiting Glut4 glucose transporters from an intracellular pool to plasma membrane (PM). To localize intracellular translocating Glut4, we studied the effects of brefeldin A (BFA), which disassembles Golgi and prevents trans-Golgi vesicular budding, on the glucose transport system. Isolated rat adipocytes were treated with and ... More
Cloning and characterization of beta-COP from Dictyostelium discoideum.
AuthorsMohrs MR, Janssen KP, Kreis T, Noegel AA, Schleicher M
JournalEur J Cell Biol
PubMed ID10887966
'We have isolated a cDNA coding for beta-COP from Dictyostelium discoideum by polymerase chain reaction using degenerate primers derived from rat beta-COP. The complete cDNA clone has a size of 2.8 kb and codes for a protein with a calculated molecular mass of 102 kDa. Dictyostelium beta-COP exhibits highest homology ... More
Association of stomatin with lipid bodies.
AuthorsUmlauf E, Csaszar E, Moertelmaier M, Schuetz GJ, Parton RG, Prohaska R
JournalJ Biol Chem
PubMed ID15024010
'The oligomeric lipid raft-associated integral protein stomatin normally localizes to the plasma membrane and the late endosomal compartment. Similar to the caveolins, it is targeted to lipid bodies (LBs) on overexpression. Endogenous stomatin also associates with LBs to a small extent. Green fluorescent protein-tagged stomatin (StomGFP) and the dominant-negative caveolin-3 ... More
A pivotal role of Rho GTPase in the regulation of morphology and function of dendritic cells.
AuthorsKobayashi M, Azuma E, Ido M, Hirayama M, Jiang Q, Iwamoto S, Kumamoto T, Yamamoto H, Sakurai M, Komada Y
JournalJ Immunol
PubMed ID11564770
'Dendritic cell (DC) is the most potent activator of CD4+ T cells and has unique dendrites and veils. To explore the function of Rho in DC, exoenzyme C3 from Clostridium botulinum was used as a specific inhibitor of Rho. Treatment of DC with C3 (DC/C3) resulted in profound morphological changes ... More
Direct measurement of endosomal pH in living cells of the rat yolk sac epithelium by laser confocal microscopy.
AuthorsIchimura T, Hatae T, Ishida T
JournalEur J Cell Biol
PubMed ID9309389
'Endocytosis of dual-fluorescent dextran by epithelial cells was observed in the perfused rat yolk sac using a confocal laser scanning microscope. Endosomal pH was quantified from confocal images using a dual-fluorescence ratiometry technique. Changes in endosomal pH were followed over time up to 32 min with and without Brefeldin A ... More
Brefeldin A, a drug that blocks secretion, prevents the assembly of non-clathrin-coated buds on Golgi cisternae.
AuthorsOrci L, Tagaya M, Amherdt M, Perrelet A, Donaldson JG, Lippincott-Schwartz J, Klausner RD, Rothman JE
JournalCell
PubMed ID2004424
'We report that brefeldin A prevents the assembly of non-clathrin-coated vesicles from Golgi cisternae in a cell-free system. This finding provides a simple molecular explanation for the primary effect of this remarkable compound in blocking constitutive secretion. We further report that when coated vesicle assembly is blocked, extensive tubule networks ... More
Identification of a JAK2-independent pathway regulating growth hormone (GH)-stimulated p44/42 mitogen-activated protein kinase activity. GH activation of Ral and phospholipase D is Src-dependent.
AuthorsZhu T, Ling L, Lobie PE
JournalJ Biol Chem
PubMed ID12218045
'We have demonstrated here that growth hormone (GH) stimulates the formation of the active GTP-bound form of both RalA and RalB in NIH-3T3 cells. Full activation of RalA and RalB by GH required the combined activity of c-Src and JAK2, both kinases activated by GH independent of the other. Activation ... More
The ARF1 GTPase-activating protein: zinc finger motif and Golgi complex localization.
AuthorsCukierman E, Huber I, Rotman M, Cassel D
JournalScience
PubMed ID8533093
'Hydrolysis of guanosine triphosphate (GTP) by the small guanosine triphosphatase (GTPase) adenosine diphosphate ribosylation factor-1 (ARF1) depends on a GTPase-activating protein (GAP). A complementary DNA encoding the ARF1 GAP was cloned from rat liver and predicts a protein with a zinc finger motif near the amino terminus. The GAP function ... More
Transport and internal organization of membranes: vesicles, membrane networks and GTP-binding proteins.
AuthorsAyala J
JournalJ Cell Sci
PubMed ID8056835
Multiple targets for brefeldin A.
AuthorsPelham HR
JournalCell
PubMed ID1934055
Brefeldin A: insights into the control of membrane traffic and organelle structure.
Brefeldin A affects the cellular distribution of endocytic receptors differentially.
AuthorsDamke H, Klumperman J, von Figura K, Braulke T
JournalBiochem Biophys Res Commun
PubMed ID1319146
The cell surface expression of three endocytic receptors was studied in human hepatoma Hep G2 cells treated with brefeldin A (BFA). Ligand binding and cell surface iodination revealed that BFA increased the number of mannose 6-phosphate/insulin-like growth factor II receptors twofold and decreased the amount of asialoglycoprotein and transferrin receptors ... More
Binding of ARF and beta-COP to Golgi membranes: possible regulation by a trimeric G protein.
The binding of cytosolic coat proteins to organelles may regulate membrane structure and traffic. Evidence is presented that a small guanosine triphosphate (GTP)-binding protein, the adenosine diphosphate ribosylation factor (ARF), reversibly associates with the Golgi apparatus in an energy, GTP, and fungal metabolite brefeldin A (BFA)-sensitive manner similar to, but ... More
An acidic motif retains vesicular monoamine transporter 2 on large dense core vesicles.
AuthorsWaites CL, Mehta A, Tan PK, Thomas G, Edwards RH, Krantz DE
JournalJ Cell Biol
PubMed ID11257117
The release of biogenic amines from large dense core vesicles (LDCVs) depends on localization of the vesicular monoamine transporter VMAT2 to LDCVs. We now find that a cluster of acidic residues including two serines phosphorylated by casein kinase 2 is required for the localization of VMAT2 to LDCVs. Deletion of ... More
Endomembrane trafficking of ras: the CAAX motif targets proteins to the ER and Golgi.
AuthorsChoy E, Chiu VK, Silletti J, Feoktistov M, Morimoto T, Michaelson D, Ivanov IE, Philips MR
JournalCell
PubMed ID10412982
We show that Nras is transiently localized in the Golgi prior to the plasma membrane (PM). Moreover, green fluorescent protein (GFP)-tagged Nras illuminated motile, peri-Golgi vesicles, and prolonged BFA treatment blocked PM expression. GFP-Hras colocalized with GFP-Nras, but GFP-Kras4B revealed less Golgi and no vesicular fluorescence. Whereas a secondary membrane ... More
ARAP1: a point of convergence for Arf and Rho signaling.
AuthorsMiura K, Jacques KM, Stauffer S, Kubosaki A, Zhu K, Hirsch DS, Resau J, Zheng Y, Randazzo PA
JournalMol Cell
PubMed ID11804590
We have identified ARAP1 and ARAP2 and examined ARAP1 as a possible link between phosphoinositide-, Arf-, and Rho-mediated cell signaling. ARAP1 contains Arf GAP, Rho GAP, Ankyrin repeat, Ras-associating, and five PH domains. In vitro, ARAP1 had Rho GAP and phosphatidylinositol (3,4,5) trisphosphate (PIP3)-dependent Arf GAP activity. ARAP1 associated with ... 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
The Golgi-associated hook3 protein is a member of a novel family of microtubule-binding proteins.
AuthorsWalenta JH, Didier AJ, Liu X, Krämer H
JournalJ Cell Biol
PubMed ID11238449
Microtubules are central to the spatial organization of diverse membrane-trafficking systems. Here, we report that Hook proteins constitute a novel family of cytosolic coiled coil proteins that bind to organelles and to microtubules. The conserved NH(2)-terminal domains of Hook proteins mediate attachment to microtubules, whereas the more divergent COOH-terminal domains ... More
Posttranscriptional gene regulation of IL-17 by the RNA-binding protein HuR is required for initiation of experimental autoimmune encephalomyelitis.
AuthorsChen J, Cascio J, Magee JD, Techasintana P, Gubin MM, Dahm GM, Calaluce R, Yu S, Atasoy U,
Journal
PubMed ID24166976
IL-17 is a proinflammatory cytokine produced by activated Th17 cells and other immune cells. IL-17-producing Th17 cells are major contributors to chronic inflammatory and autoimmune diseases, such as multiple sclerosis, rheumatoid arthritis, and inflammatory bowel disease. Although the transcriptional regulation of Th17 cells is well understood, the posttranscriptional regulation of ... More
CMV-specific CD8+ T-cell function is not impaired in chronic lymphocytic leukemia.
Authorste Raa GD, Pascutti MF, García-Vallejo JJ, Reinen E, Remmerswaal EB, ten Berge IJ, van Lier RA, Eldering E, van Oers MH, Tonino SH, Kater AP,
Journal
PubMed ID24246502
In chronic lymphocytic leukemia (CLL), CD8(+) T cells exhibit features of exhaustion and impaired functionality. Yet, reactivations of latent viruses such as cytomegalovirus (CMV) are uncommon in untreated CLL, suggesting that antiviral responses are uncompromised. We analyzed phenotypical and functional characteristics of CMV-specific CD8(+) T cells in CLL patients in ... More
Diverse subcellular locations of cryptogein-induced reactive oxygen species production in tobacco Bright Yellow-2 cells.
AuthorsAshtamker C, Kiss V, Sagi M, Davydov O, Fluhr R,
JournalPlant Physiol
PubMed ID17277088
Reactive oxygen species (ROS) play a crucial role in many cellular responses and signaling pathways, including the oxidative burst defense response to pathogens. We have examined very early events in cryptogein-induced ROS production in tobacco (Nicotiana tabacum) Bright Yellow-2 suspension cells. Using Amplex Red and Amplex Ultra Red reagents, which ... More
Matrix proteins can generate the higher order architecture of the Golgi apparatus.
AuthorsSeemann J, Jokitalo E, Pypaert M, Warren G
JournalNature
PubMed ID11069184
The Golgi apparatus in animal cells comprises a reticulum of linked stacks in the pericentriolar and often in the juxtanuclear regions of the cell. The unique architecture of this organelle is thought to depend on the cytoskeleton and cytoplasmic matrix proteins--the best characterized being the golgin family of fibrous, coiled-coil ... More
Receptor and protein kinase C-mediated regulation of ARF binding to the Golgi complex.
AuthorsDe Matteis MA, Santini G, Kahn RA, Di Tullio G, Luini A
JournalNature
PubMed ID7689177
The formation of constitutive transport vesicles involves the association of non-clathrin coat proteins to transport organelles. Here we report that IgE receptors and protein kinase C (PKC) regulate the GTP-dependent binding of the two coat proteins ADP-ribosylation factor (ARF) and beta-COP to Golgi membranes in rat basophilic leukaemia cells. Activation ... More
Rapid redistribution of Golgi proteins into the ER in cells treated with brefeldin A: evidence for membrane cycling from Golgi to ER.
In cells treated with brefeldin A (BFA), movement of newly synthesized membrane proteins from the endoplasmic reticulum (ER) to the Golgi apparatus was blocked. Surprisingly, the glycoproteins retained in the ER were rapidly processed by cis/medial Golgi enzymes but not by trans Golgi enzymes. An explanation for these observations was ... More
Golgi membranes are absorbed into and reemerge from the ER during mitosis.
AuthorsZaal KJ, Smith CL, Polishchuk RS, Altan N, Cole NB, Ellenberg J, Hirschberg K, Presley JF, Roberts TH, Siggia E, Phair RD, Lippincott-Schwartz J
JournalCell
PubMed ID10612395
Quantitative imaging and photobleaching were used to measure ER/Golgi recycling of GFP-tagged Golgi proteins in interphase cells and to monitor the dissolution and reformation of the Golgi during mitosis. In interphase, recycling occurred every 1.5 hr, and blocking ER egress trapped cycling Golgi enzymes in the ER with loss of ... More
Brefeldin A-induced increase of sphingomyelin synthesis. Assay for the action of the antibiotic in mammalian cells.
AuthorsBrüning A, Karrenbauer A, Schnabel E, Wieland FT
JournalJ Biol Chem
PubMed ID1544889
Brefeldin A leads to an increase of sphingomyelin in Chinese hamster ovary cells. The antibiotic is known to cause a dramatic morphological change of the endomembrane system in various mammalian cells resulting in a redistribution of Golgi resident proteins to the endoplasmic reticulum (Lippincott-Schwartz, J., Donaldson, J. G., Schweizer, A., ... More
Synaptotagmin IV is present at the Golgi and distal parts of neurites.
AuthorsIbata K, Fukuda M, Hamada T, Kabayama H, Mikoshiba K
JournalJ Neurochem
PubMed ID10646502
Synaptotagmin IV (SytIV) is an immediate early gene induced by membrane depolarization in PC12 cells and in rat brain. However, little is known about the function of SytIV or the functional relationship between SytIV and SytI, because SytIV has yet to be localized. Here we show that SytIV was localized ... More
Disruption of the Golgi apparatus by brefeldin A inhibits the cytotoxicity of ricin, modeccin, and Pseudomonas toxin.
AuthorsYoshida T, Chen CC, Zhang MS, Wu HC
JournalExp Cell Res
PubMed ID1899070
We have studied the cytotoxicity of ricin in cells treated with brefeldin A (BFA), which dramatically disrupts the structure of the Golgi apparatus causing Golgi content and membrane to redistribute to the ER. BFA inhibits the cytotoxicity of ricin in Chinese hamster ovary, normal rat kidney, and Vero cells and ... 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
Trax (translin-associated factor X), a primarily cytoplasmic protein, inhibits the binding of TB-RBP (translin) to RNA.
AuthorsChennathukuzhi VM, Kurihara Y, Bray JD, Hecht NB
JournalJ Biol Chem
PubMed ID11278549
Trax (Translin-associated factor X) has been shown to interact with TB-RBP/Translin by its coimmunoprecipitation and in yeast two-hybrid assays. Here we demonstrate that Trax is widely expressed, does not bind to DNA or RNA, but forms heterodimers with TB-RBP under reducing conditions. The heterodimer of TB-RBP and Trax inhibits TB-RBP ... More
Rapid turnover of low-density lipoprotein receptor by a non-lysosomal pathway in mouse macrophage J774 cells and inhibitory effect of brefeldin A.
AuthorsShite S, Seguchi T, Shimada T, Ono M, Kuwano M
JournalEur J Biochem
PubMed ID2384096
The low-density lipoprotein (LDL) receptor of molecular mass 155 kDa was expressed on the cell surface of cultured mouse macrophage J774 cells. The conversion rate of precursor to mature form of LDL receptor in J774 cells was comparable to that in mouse fibroblast L cells. The half-life of the LDL ... More
Transport of glutathione conjugates into secretory vesicles is mediated by the multidrug-resistance protein 1.
AuthorsVan Luyn MJ, Müller M, Renes J, Meijer C, Scheper RJ, Nienhuis EF, Mulder NH, Jansen PL, De Vries EG
JournalInt J Cancer
PubMed ID9533762
Intracellular glutathione-conjugate transport was evaluated in the human small cell lung carcinoma cell line GLC4 with low multidrug resistance protein (MRP1) expression and its 300x doxorubicin-resistant, MRP1-over-expressing, GLC4-Adr subline. Transport of non-toxic concentrations of monochlorobimane and 5-chloro-methyl fluorescein diacetate was evaluated using fluorescence microscopy. After exposure to these compounds, fluorescence ... More
Endocytosed transferrin receptors recycle via distinct dynamin and phosphatidylinositol 3-kinase-dependent pathways.
Authorsvan Dam EM, Ten Broeke T, Jansen K, Spijkers P, Stoorvogel W
JournalJ Biol Chem
PubMed ID12372835
Recycling of endocytosed membrane proteins involves passage through early endosomes and recycling endosomes. Previously, we demonstrated a role for clathrin-coated vesicles in transferrin receptor recycling. These clathrin-coated vesicles are formed from recycling endosomes in a process that was inhibited in dynamin-1(G273D)-overexpressing cells. Here we show a second transferrin recycling pathway, ... More
Brefeldin A as a regulator of grp78 gene expression in mammalian cells.
AuthorsLiu ES, Ou JH, Lee AS
JournalJ Biol Chem
PubMed ID1551919
We report here that brefeldin A (BFA), which specifically blocks protein transport from an endoplasmic reticulum to the Golgi apparatus and causes resorption of Golgi membrane to the endoplasmic reticulum, specifically induced the endoplasmic reticulum-resident protein GRP78. Treatment of a human hepatoma cell line Alex-PC with BFA at a concentration ... More
Uptake and intracellular transport of acidic fibroblast growth factor: evidence for free and cytoskeleton-anchored fibroblast growth factor receptors.
AuthorsCitores L, Wesche J, Kolpakova E, Olsnes S
JournalMol Biol Cell
PubMed ID10564275
Endocytic uptake and intracellular transport of acidic FGF was studied in cells transfected with FGF receptor 4 (FGFR4). Acidification of the cytosol to block endocytic uptake from coated pits did not inhibit endocytosis of the growth factor in COS cells transfected with FGFR4, indicating that it is to a large ... More
Brefeldin A-dependent membrane tubule formation reconstituted in vitro is driven by a cell cycle-regulated microtubule motor.
AuthorsRobertson AM, Allan VJ
JournalMol Biol Cell
PubMed ID10712511
Treatment of cultured cells with brefeldin A (BFA) induces the formation of extensive membrane tubules from the Golgi apparatus, trans-Golgi network, and early endosomes in a microtubule-dependent manner. We have reconstituted this transport process in vitro using Xenopus egg cytosol and a rat liver Golgi-enriched membrane fraction. The presence of ... More
Accumulation of caveolin in the endoplasmic reticulum redirects the protein to lipid storage droplets.
AuthorsOstermeyer AG, Paci JM, Zeng Y, Lublin DM, Munro S, Brown DA
JournalJ Cell Biol
PubMed ID11238461
Caveolin-1 is normally localized in plasma membrane caveolae and the Golgi apparatus in mammalian cells. We found three treatments that redirected the protein to lipid storage droplets, identified by staining with the lipophilic dye Nile red and the marker protein ADRP. Caveolin-1 was targeted to the droplets when linked to ... More
Differential survival of transferred CD8 T cells and host reconstitution depending on TCR avidity for host-expressed alloantigen.
AuthorsAuphan-Anezin N, Schmitt-Verhulst AM
JournalJ Immunol
PubMed ID11390468
We transferred naive alloreactive CD8 T cells from TCR transgenic mice to irradiated recipients expressing a partial (H-2Kbm8) or a full (H-2Kb) agonist alloantigen (alloAg). The consequences were strikingly distinct, resulting in acceleration of host lymphopoiesis in the former group, but in strong graft-vs-host reaction, preventing host lymphocyte reconstitution in ... More
A prion protein cycles between the cell surface and an endocytic compartment in cultured neuroblastoma cells.
AuthorsShyng SL, Huber MT, Harris DA
JournalJ Biol Chem
PubMed ID8101844
The prion protein (PrPC) is a glycolipid-anchored, cell surface protein of unknown function, a posttranslationally modified isoform of which has been implicated in the pathogenesis of spongiform encephalopathies in man and animals. We report here the novel observation that chPrP, the chicken homologue of mammalian PrPC, constitutively cycles between the ... More
Okadaic acid treatment leads to a fragmentation of the trans-Golgi network and an increase in expression of TGN38 at the cell surface.
AuthorsHorn M, Banting G
JournalBiochem J
PubMed ID8037693
Okadaic acid (OA) is a protein phosphatase inhibitor which has, among other properties, previously been shown to induce a fragmentation of the cisternae of the Golgi stack [for review, see Lucocq (1992) J. Cell Sci. 103, 875-880]. The effects of OA an reversible and mimic intracellular events which occur during ... More
Quality control in the endoplasmic reticulum: folding and misfolding of vesicular stomatitis virus G protein in cells and in vitro.
Authorsde Silva AM, Balch WE, Helenius A
JournalJ Cell Biol
PubMed ID1697299
Parallel experiments in living cells and in vitro were undertaken to characterize the mechanism by which misfolded and unassembled glycoproteins are retained in the ER. A thermoreversible folding mutant of vesicular stomatitis virus (VSV) G protein called ts045 was analyzed. At 39 degrees C, newly synthesized G failed to fold ... More
Mycobacterium avium- and Mycobacterium tuberculosis-containing vacuoles are dynamic, fusion-competent vesicles that are accessible to glycosphingolipids from the host cell plasmalemma.
AuthorsRussell DG, Dant J, Sturgill-Koszycki S
JournalJ Immunol
PubMed ID8648123
The vacuoles inhabited by viable Mycobacterium avium and Mycobacterium tuberculosis show limited fusion with endosomal and lysosomal compartments. This ability to regulate the maturation of their phagosomal compartments and restrict their differentiation into hydrolytically active vacuoles appears to correlate with the survival of the bacilli. Data presented in this current ... More
Fluorescent conjugates of brefeldin A selectively stain the endoplasmic reticulum and Golgi complex of living cells.
AuthorsDeng Y, Bennink JR, Kang HC, Haugland RP, Yewdell JW
JournalJ Histochem Cytochem
PubMed ID7543914
The fungal metabolite brefeldin A (BFA) interferes with vesicular trafficking in most animal cells. To gain insight into the mechanism of BFA action, we esterified it to the fluorophore, boron dipyromethene difluoride (BODIPY). BODIPY-BEA localized predominantly in the endoplasmic reticulum (ER) and Golgi complex of viable cells and was extracted ... More
A requirement for ARF6 in Fcgamma receptor-mediated phagocytosis in macrophages.
AuthorsZhang Q, Cox D, Tseng CC, Donaldson JG, Greenberg S
JournalJ Biol Chem
PubMed ID9685333
Phagocytosis requires extension of F-actin-rich pseudopods and is accompanied by membrane fusion events. Members of the ARF family of GTPases are essential for many aspects of membrane trafficking. To test a role for this family of proteins in Fcgamma receptor-mediated phagocytosis, we utilized the fungal metabolite brefeldin A (BFA). The ... More
EEA1, a tethering protein of the early sorting endosome, shows a polarized distribution in hippocampal neurons, epithelial cells, and fibroblasts.
AuthorsWilson JM, de Hoop M, Zorzi N, Toh BH, Dotti CG, Parton RG
JournalMol Biol Cell
PubMed ID10930461
EEA1 is an early endosomal Rab5 effector protein that has been implicated in the docking of incoming endocytic vesicles before fusion with early endosomes. Because of the presence of complex endosomal pathways in polarized and nonpolarized cells, we have examined the distribution of EEA1 in diverse cell types. Ultrastructural analysis ... More
Flotillin-1/reggie-2 traffics to surface raft domains via a novel golgi-independent pathway. Identification of a novel membrane targeting domain and a role for palmitoylation.
AuthorsMorrow IC, Rea S, Martin S, Prior IA, Prohaska R, Hancock JF, James DE, Parton RG
JournalJ Biol Chem
PubMed ID12370178
Flotillins are lipid raft-associated proteins, which have been implicated in neuronal regeneration and insulin signaling. We now show that newly synthesized flotillin-1 reaches the plasma membrane via a Sar1-independent and brefeldin A-resistant targeting pathway. Consistent with post-translational membrane association of flotillin, protease sensitivity experiments suggest that flotillin-1 is not a ... More
Translocation of oxysterol binding protein to Golgi apparatus triggered by ligand binding.
AuthorsRidgway ND, Dawson PA, Ho YK, Brown MS, Goldstein JL
JournalJ Cell Biol
PubMed ID1730758
A cDNA encoding a cytoplasmic oxysterol binding protein was expressed at high levels by transfection in animal cells. This protein binds oxysterols such as 25-hydroxycholesterol that regulate sterol metabolism by transcriptional and posttranscriptional effects. In the transfected cells, some of the oxysterol binding protein (OSBP) was distributed diffusely in the ... 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
Lack of palmitoylation redirects p59Hck from the plasma membrane to p61Hck-positive lysosomes.
AuthorsCarréno S, Gouze ME, Schaak S, Emorine LJ, Maridonneau-Parini I
JournalJ Biol Chem
PubMed ID10967098
Hck, a protein-tyrosine kinase of phagocytes, is the unique member of the Src family expressed under two alternatively translated isoforms differing in their N-terminal site of acylation: p61(Hck) has an additional 21-amino acid sequence comprising a single myristoylation motif, whereas p59(Hck) N terminus has myristoylation and palmitoylation sites. To identify ... More
We examined the effect of leptin on renal function and renal Na(+),K(+)-ATPase and ouabain-sensitive H(+),K(+)-ATPase activities in the rat. Leptin was infused under general anaesthesia into the abdominal aorta proximally to the renal arteries. Leptin infused at doses of 1 and 10 microg/kg/min increased urine output by 40% and 140%, ... More
Stage-specific expression and targeting of cyst wall protein-green fluorescent protein chimeras in Giardia.
AuthorsHehl AB, Marti M, Köhler P
JournalMol Biol Cell
PubMed ID10793152
In preparation for being shed into the environment as infectious cysts, trophozoites of Giardia spp. synthesize and deposit large amounts of extracellular matrix into a resistant extracellular cyst wall. Functional aspects of this developmentally regulated process were investigated by expressing a series of chimeric cyst wall protein 1 (CWP1)-green fluorescent ... More
Inhibition by brefeldin A of presentation of exogenous protein antigens to MHC class II-restricted T cells.
AuthorsAdorini L, Ullrich SJ, Appella E, Fuchs S
JournalNature
PubMed ID2366863
Peptides bound to class I or class II major histocompatibility complex (MHC)-encoded molecules are ligands for the antigen-specific T-cell receptor of T-cells carrying the CD8 and CD4 antigens, respectively. MHC class I-restricted T cells generally recognize peptides derived from processing of endogenously synthesized cellular antigens, whereas class II-restricted T cells ... More
Perturbation of the morphology of the trans-Golgi network following Brefeldin A treatment: redistribution of a TGN-specific integral membrane protein, TGN38.
AuthorsReaves B, Banting G
JournalJ Cell Biol
PubMed ID1730751
Brefeldin A (BFA) has a dramatic effect on the morphology of the Golgi apparatus and induces a rapid redistribution of Golgi proteins into the ER (Lippincott-Schwartz, J., L. C. Yuan, J. S. Bonifacino, and R. D. Klausner. 1989. Cell. 56:801-813). To date, no evidence that BFA affects the morphology of ... More
Reconstitution of brefeldin A-induced golgi tubulation and fusion with the endoplasmic reticulum in semi-intact chinese hamster ovary cells.
AuthorsKano F, Sako Y, Tagaya M, Yanagida T, Murata M
JournalMol Biol Cell
PubMed ID10982401
The fungal metabolite brefeldin A (BFA) induces the disassembly of the Golgi complex in mammalian cells. The drug seems to accentuate tubule formation and causes the subsequent fusion with the endoplasmic reticulum (ER). To investigate the biochemical requirements and kinetics of BFA-induced Golgi disassembly, we have reconstituted the process of ... More
Dissociation of a 110-kD peripheral membrane protein from the Golgi apparatus is an early event in brefeldin A action.
AuthorsDonaldson JG, Lippincott-Schwartz J, Bloom GS, Kreis TE, Klausner RD
JournalJ Cell Biol
PubMed ID2277061
Brefeldin A (BFA) has a profound effect on the structure of the Golgi apparatus, causing Golgi proteins to redistribute into the ER minutes after drug treatment. Here we describe the dissociation of a 110-kD cytoplasmically oriented peripheral membrane protein (Allan, V. J., and T. E. Kreis. 1986. J. Cell Biol. ... More