DOCSLIB.ORG

Three Ways to Make a Vesicle

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Three Ways to Make a Vesicle REVIEWS THREE WAYS TO MAKE A VESICLE Tomas Kirchhausen Cargo molecules have to be included in carrier vesicles of different forms and sizes to be transported between organelles. During this process, a limited set of proteins, including the coat proteins COPI, COPII and clathrin, carries out a programmed set of sequential interactions that lead to the budding of vesicles. A general model to explain the formation of coated vesicles is starting to emerge but the picture is more complex than we had imagined. ENDOCYTIC PATHWAY Transport of proteins and lipids along the ENDOCYTIC or energy dependent and includes sorting of cargo to the Macromolecules are taken up SECRETORY PATHWAYS is a hallmark of eukaryotic cells. The forming coat. Coat propagation, the second step in the by invagination of the plasma membrane fluxes along these pathways are very large process, couples further addition of coat components membrane. They first arrive in and rapid. A FIBROBLAST kept in resting conditions in a and additional recruitment of cargo with invagination of early endosomes, then late endosomes, and finally tissue culture plate internalizes an amount of mem- the underlying membrane. When formation of the coat lysosomes, where they are brane equivalent to the whole surface area of the cell in ends, the vesicle buds by scission of the neck connecting degraded by hydrolases. one hour. Inside the cell, it often takes only seconds for a the deeply invaginated membrane to the donor surface. carrier vesicle to move from the donor membrane to This is a relatively simple step for COPI- and COPII- SECRETORY PATHWAY the acceptor organelle. But intracellular traffic is not coated vesicles but involves a significant amount of regu- Secretory or membrane proteins are inserted into the only rapid, it is also very selective. Only a subset of the lation for clathrin-coated vesicles. Finally, during uncoat- endoplasmic reticulum. They proteins and lipids in the donor membrane are allowed ing, the coat components are released so that membrane are then transported through into the transport vesicle, effectively preventing the fusion can occur between the naked vesicle and the target the Golgi to the trans-Golgi homogenization of membrane components and per- organelle. This article discusses the emerging molecular network, where they are sorted to their final destination. mitting membranous organelles to maintain distinct rationale for coated-vesicle assembly for, in order of identities throughout the life of the cell. increasing complexity, COPII, COPI and clathrin. FIBROBLAST Considerable progress has been made towards under- Common cell type found in standing the molecular basis of membrane traffic1–8.A Pathways of membrane traffic connective tissue in many parts number of traffic pathways have been defined, their COPI and COPII vesicles traffic between the endoplas- of the body, which secretes an extracellular matrix rich in major protein components identified and the structures mic reticulum (ER) and the Golgi complex — COPI collagen and other of several of the key components determined at atomic primarily from the Golgi to the ER and between Golgi macromolecules and connects or molecular resolution. The best studied traffic path- cisternae, and COPII from the ER to the Golgi (FIG. 1). cell layers. ways are those that use carrier vesicles that are clearly The clathrin pathway has two major routes, from the MACROPINOCYTOSIS identifiable by their coats, made of the coatomer COPI, plasma membrane to the early endosome and from the Actin-dependent process by of COPII, or of clathrin and its partners (FIG. 1).During Golgi to the endosome. Other structures have been which cells engulf large volumes the formation of a vesicle, a limited set of coat proteins observed in the cell that do not have COP or clathrin of fluids. (TABLE 1) carries out a programmed set of sequential coats. Internalization from the plasma membrane can interactions that lead to budding from the parent mem- also occur via MACROPINOCYTOSIS, PHAGOCYTOSIS and proba- brane, uncoating, fusion with a target membrane and bly through CAVEOLAE9. In the secretory pathway, poorly recycling of the coat components. There are clear similar- understood tube-like structures connect the Golgi with Harvard Medical School, ities and differences between the ways that COPI, COPII the plasma membrane and the ER10–13. In the endocytic 200 Longwood Avenue, and clathrin coats handle these steps (TABLE 2). pathway, tubules emerge from early endosomes and Boston, Massachusetts 14 02115, USA. e-mail: The first group of reactions, leading to the specific participate in recycling to the plasma membrane . kirchhausen@crystal. recruitment of coat components to the corresponding These carrier structures have been hard to study, partly harvard.edu donor membrane, form the initiation step. This step is because they tend to be heterogeneous. In some cases, NATURE REVIEWS | MOLECULAR CELL BIOLOGY © 2000 Macmillan Magazines Ltd VOLUME 1 | DECEMBER 2000 | 187 REVIEWS molecular detail. Nevertheless, there is good agreement Clathrin coats in the field that the major components have been identi- Recycling endosome fied and there is a good working model for the sequence in which they act (FIG. 2). Biochemical and cell biological observations. Although several as-yet-undefined membrane components are Early endosome Late endosome presumably necessary for efficient operation of the COPII pathway, vesicular transport can be minimally Clathrin coats reconstituted using three cytosolic components contain- ing a total of five proteins: the Sec23p–Sec24p complex, the Sec13p–Sec31p complex and the small GTPase Trans-Golgi Sar1p (REF.16). These proteins support a cargo-carrying budding reaction from isolated ER membranes. Deletion of the genes for most of these components is COPII coats Cis-Golgi lethal for yeast, highlighting the strict dependence on COPI coats this pathway for ER-to-Golgi traffic. Lysosome The events driven by these five cytosolic proteins, Endoplasmic reticulum (ER) together with the membrane components required for targeting and fusion, are varied and complex (TABLE 2). Cargo capture, deformation of the budding membrane, scission to detach the forming vesicle from the donor membrane and coat release before fusion of the vesicle with the Golgi network all occur in this minimal system. Figure 1 | The major membrane traffic pathways that use carrier vesicles coated with The GTP-binding protein Sar1p is particularly impor- COPI, COPII and clathrin in eukaryotic cells. In the biosynthetic pathway, newly tant for the budding reaction because its activation initi- synthesized molecules are transported from the endoplasmic reticulum to the Golgi and from ates coat formation; it also recruits part of the ‘label’ one cisterna of the Golgi to the next until they reach the trans-Golgi network. There, sorting needed for correct vesicle targeting and fusion. The occurs, directing traffic to the plasma membrane or to endosomes. In the endocytic pathway, GDP-bound form of Sar1p is normally cytosolic and is macromolecules are internalized at the plasma membrane and forwarded to early endosomes, from where they are either recycled to the plasma membrane through recycling endosomes or recruited to the ER membrane upon interaction with forwarded towards degradation in late endosomes and lysosomes. Sec12p, an ER-bound membrane protein that serves as the guanine exchange factor (GEF) for Sar1p (REF.17). Sar1p–GTP then facilitates the association of the they might be used for large-scale movement of selec- Sec23p–Sec24p complex with cargo proteins. tively captured membrane components12,15. The most The Sec23p–Sec24p complex is probably the com- important distinction between coated vesicles and other ponent responsible for cargo recognition18,19 but the forms of membrane carriers is the presence of an iden- sorting signals recognized by the complex remain to be tifiable protein coat, which assembles at a particular identified. Members of the p24 family of transmem- region of the membrane, locally deforming it. Although brane proteins bind to Sec23p through a cytosolic the events required are similar — tubes, diphenylalanine motif. As these proteins are required macropinosomes and phagosomes also need to form, for efficient ER-to-Golgi traffic of some cargo pinch off, reach their target and fuse with it just like proteins20, it is thought that they might serve as cargo coated vesicles — it is impossible to transfer our under- adaptors21. In addition to recruiting the Sec23p–Sec24p standing of vesicle traffic to these other mechanisms. complex, the GTP-bound form of Sar1p activates Sec23p to bind SNARE proteins involved in the speci- COPII ficity of targeting and in the fusion reaction of vesicles COPII components, and COPII-coated vesicles, were with acceptor membranes18. ER membranes with originally discovered in the yeast Saccharomyces cerevisiae Sec23p–Sec24p and Sar1p can then recruit using genetic approaches coupled with a cell-free assay Sec13p–Sec31p (REF.16). The complex is likely to act as a that measured the transfer of a marker protein from the scaffold, very much like clathrin, to drive membrane 3,8 PHAGOCYTOSIS ER to the Golgi . This pathway has a mammalian coun- deformation and to complete vesicle budding. Actin-dependent process, by terpart and most of the COPII components have been Completing the cycle, Sec23p acts as a GTPase-activat- which cells engulf external identified (TABLE 1). Newly synthesized proteins destined ing protein (GAP) for Sar1p. It is thought that, after particulate material by for secretion are sorted into COPII-coated vesicles at spe- GTP hydrolysis, Sar1p–GDP is released, leading to extension and fusion of pseudopods around each cialized regions of the ER, which in mammalian cells do uncoating before fusion of the vesicle to the target particle. not contain membrane-bound ribosomes. membrane and formation of a new coated vesicle. GTP Biochemical analysis has led to a relatively detailed hydrolysis by Sar1p is thus a timer, triggering uncoating CAVEOLA model for the mechanism of COPII vesicle formation. at a suitable interval after coat formation.
Load more

Recommended publications
  • Mea6 Controls VLDL Transport Through the Coordinated Regulation of COPII Assembly
    Cell Research (2016) 26:787-804. npg © 2016 IBCB, SIBS, CAS All rights reserved 1001-0602/16 $ 32.00 ORIGINAL ARTICLE www.nature.com/cr Mea6 controls VLDL transport through the coordinated regulation of COPII assembly Yaqing Wang1, *, Liang Liu1, 2, *, Hongsheng Zhang1, 2, Junwan Fan1, 2, Feng Zhang1, 2, Mei Yu3, Lei Shi1, Lin Yang1, Sin Man Lam1, Huimin Wang4, Xiaowei Chen4, Yingchun Wang1, Fei Gao5, Guanghou Shui1, Zhiheng Xu1, 6 1State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China; 2University of Chinese Academy of Sciences, Beijing 100101, China; 3School of Life Science, Shandong University, Jinan 250100, China; 4Institute of Molecular Medicine, Peking University, Beijing 100871, China; 5State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China; 6Translation- al Medical Center for Stem Cell Therapy, Shanghai East Hospital, Tongji University School of Medicine, Shanghai 200120, China Lipid accumulation, which may be caused by the disturbance in very low density lipoprotein (VLDL) secretion in the liver, can lead to fatty liver disease. VLDL is synthesized in endoplasmic reticulum (ER) and transported to Golgi apparatus for secretion into plasma. However, the underlying molecular mechanism for VLDL transport is still poor- ly understood. Here we show that hepatocyte-specific deletion of meningioma-expressed antigen 6 (Mea6)/cutaneous T cell lymphoma-associated antigen 5C (cTAGE5C) leads to severe fatty liver and hypolipemia in mice. Quantitative lip- idomic and proteomic analyses indicate that Mea6/cTAGE5 deletion impairs the secretion of different types of lipids and proteins, including VLDL, from the liver.
    [Show full text]
  • Clathrin-Independent Pathways of Endocytosis
    Downloaded from http://cshperspectives.cshlp.org/ on October 3, 2021 - Published by Cold Spring Harbor Laboratory Press Clathrin-Independent Pathways of Endocytosis Satyajit Mayor1, Robert G. Parton2, and Julie G. Donaldson3 1National Centre for Biological Sciences, Tata Institute of Fundamental Research, and Institute for Stem Cell Biology and Regenerative Medicine, Bangalore 560065, India 2The University of Queensland, Institute for Molecular Bioscience and Centre for Microscopy and Microanalysis, Queensland 4072, Brisbane, Australia 3Cell Biology and Physiology Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892 Correspondence: [email protected] There are many pathways of endocytosis at the cell surface that apparently operate at the same time. With the advent of new molecular genetic and imaging tools, an understanding of the different ways by which a cell may endocytose cargo is increasing by leaps and bounds. In this review we explore pathways of endocytosis that occur in the absence of clathrin. These are referred to as clathrin-independent endocytosis (CIE). Here we primarily focus on those pathways that function at the small scale in which some have distinct coats (caveolae) and others function in the absence of specific coated intermediates. We follow the trafficking itineraries of the material endocytosed by these pathways and finally discuss the functional roles that these pathways play in cell and tissue physiology. It is likely that these pathways will play key roles in the regulation of plasma membrane area and tension and also control the availability of membrane during cell migration. he identification of many of the components Consequently, CME has remained a pre- Tinvolved in clathrin-mediated endocytosis dominant paradigm for following the uptake (CME) and their subsequent characterization of material into the cell.
    [Show full text]
  • Coatomer-Rich Endoplasmic Reticulum LELIO ORCI*, ALAIN PERRELET*, MARIELLA RAVAZZOLA*, Myltne AMHERDT*, JAMES E
    Proc. Nati. Acad. Sci. USA Vol. 91, pp. 11924-11928, December 1994 Cell Biology Coatomer-rich endoplasmic reticulum LELIO ORCI*, ALAIN PERRELET*, MARIELLA RAVAZZOLA*, MYLtNE AMHERDT*, JAMES E. ROTHMANt, AND RANDY SCHEKMAN* *Department of Morphology, University of Geneva Medical School, 1211 Geneva 4, Switzerland; tDivision of Biochemistry and Molecular Biology, Howard Hughes Medical Institute, University of California, Berkeley, CA 94720; and tCellular Biochemistry and Biophysics Program, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10021 Contributed by Randy Schekman, August 30, 1994 ABSTRACT We identify in normal cells the existence of digestion and incubated at normal (370C) or low (220C, 15'C, two distinct sites of the transitional endoplasmic reticulum 40C) temperatures under continuous shaking and gassing with (ER), one housing the Sec23p protein complex (the classical 95% 02/5% CO2. Monolayer cultures ofpancreatic endocrine transitional element), the other the coatomer protein complex cells (24) were exposed to low temperatures as for isolated (the coatomer-rich ER). Experimental conditions that reduce islets. ATP depletion in both preparations was induced at transport from the ER to the Golgi complex lead to the 370C with 10 ImM antimycin or 1 mM dinitrophenol or by overexpression of this newly defined coatomer-rich ER. gassing the cells with N2 instead of the 02/CO2 mixture. At the end of the incubations, the samples were fixed with Progress in the identification and characterization of the 1% glutaraldehyde in 0.1 M sodium phosphate buffer (pH 7.4) carriers involved in membrane traffic from the endoplasmic for 1 hr at the final temperature of the incubation protocol, reticulum (ER) to and through the Golgi complex was pos- washed with buffer, and processed for Epon embedding sible by combining morphological and biochemical methods, (conventional thin sections) or for cryoultramicrotomy ac- cell-free assay systems, and yeast genetics (for review see refs.
    [Show full text]
  • An Arf1 Synthetic Lethal Screen Identifies a New Clathrin Heavy
    Copyright 1998 by the Genetics Society of America An arf1D Synthetic Lethal Screen Identi®es a New Clathrin Heavy Chain Conditional Allele That Perturbs Vacuolar Protein Transport in Saccharomyces cerevisiae Chih-Ying Chen and Todd R. Graham Department of Molecular Biology, Vanderbilt University, Nashville, Tennessee 37235 Manuscript received March 5, 1998 Accepted for publication June 16, 1998 ABSTRACT ADP-ribosylation factor (ARF) is a small GTP-binding protein that is thought to regulate the assembly of coat proteins on transport vesicles. To identify factors that functionally interact with ARF, we have performed a genetic screen in Saccharomyces cerevisiae for mutations that exhibit synthetic lethality with an arf1D allele and de®ned seven genes by complementation tests (SWA1-7 for synthetically lethal with arf1D). Most of the swa mutants exhibit phenotypes comparable to arf1D mutants such as temperature-conditional growth, hypersensitivity to ¯uoride ions, and partial protein transport and glycosylation defects. Here, we report that swa5-1 is a new temperature-sensitive allele of the clathrin heavy chain gene (chc1-5), which carries a frameshift mutation near the 39 end of the CHC1 open reading frame. This genetic interaction between arf1 and chc1 provides in vivo evidence for a role for ARF in clathrin coat assembly. Surprisingly, strains harboring chc1-5 exhibited a signi®cant defect in transport of carboxypeptidase Y or carboxypepti- dase S to the vacuole that was not observed in other chc1 ts mutants. The kinetics of invertase secretion or transport of alkaline phosphatase to the vacuole were not signi®cantly affected in the chc1-5 mutant, further implicating clathrin speci®cally in the Golgi to vacuole transport pathway for carboxypeptidase Y.
    [Show full text]
  • Formation of COPI-Coated Vesicles at a Glance Eric C
    © 2018. Published by The Company of Biologists Ltd | Journal of Cell Science (2018) 131, jcs209890. doi:10.1242/jcs.209890 CELL SCIENCE AT A GLANCE Formation of COPI-coated vesicles at a glance Eric C. Arakel1 and Blanche Schwappach1,2,* ABSTRACT unresolved, this review attempts to refocus the perspectives of The coat protein complex I (COPI) allows the precise sorting of lipids the field. and proteins between Golgi cisternae and retrieval from the Golgi KEY WORDS: Arf1, ArfGAP, COPI, Coatomer, Golgi, Endoplasmic to the ER. This essential role maintains the identity of the early reticulum, Vesicle coat secretory pathway and impinges on key cellular processes, such as protein quality control. In this Cell Science at a Glance and accompanying poster, we illustrate the different stages of COPI- Introduction coated vesicle formation and revisit decades of research in the Vesicle coat proteins, such as the archetypal clathrin and the coat context of recent advances in the elucidation of COPI coat structure. protein complexes II and I (COPII and COPI, respectively) are By calling attention to an array of questions that have remained molecular machines with two central roles: enabling vesicle formation, and selecting protein and lipid cargo to be packaged within them. Thus, coat proteins fulfil a central role in the 1Department of Molecular Biology, Universitätsmedizin Göttingen, Humboldtallee homeostasis of the cell’s endomembrane system and are the basis 23, 37073 Göttingen, Germany. 2Max-Planck Institute for Biophysical Chemistry, 37077 Göttingen, Germany. of functionally segregated compartments. COPI operates in retrieval from the Golgi to the endoplasmic reticulum (ER) and in intra-Golgi *Author for correspondence ([email protected]) transport (Beck et al., 2009; Duden, 2003; Lee et al., 2004a; Spang, E.C.A., 0000-0001-7716-7149; B.S., 0000-0003-0225-6432 2009), and maintains ER- and Golgi-resident chaperones and enzymes where they belong.
    [Show full text]
  • ADP-Ribosylation Factor, a Small GTP-Binding Protein, Is Required for Binding of the Coatomer Protein Fl-COP to Golgi Membranes JULIE G
    Proc. Natl. Acad. Sci. USA Vol. 89, pp. 6408-6412, July 1992 Biochemistry ADP-ribosylation factor, a small GTP-binding protein, is required for binding of the coatomer protein fl-COP to Golgi membranes JULIE G. DONALDSON*, DAN CASSEL*t, RICHARD A. KAHN*, AND RICHARD D. KLAUSNER* *Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, and tLaboratory of Biological Chemistry, Division of Cancer Treatment, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892 Communicated by Marc Kirschner, April 20, 1992 (receivedfor review February 11, 1992) ABSTRACT The coatomer is a cytosolic protein complex localized to the Golgi complex, although their functions have that reversibly associates with Golgi membranes and is Impli- not been defined. Distinct among these proteins is the ADP- cated in modulating Golgi membrane transport. The associa- ribosylation factor (ARF), originally identified as a cofactor tion of 13-COP, a component of coatomer, with Golgi mem- required for in vitro cholera toxin-catalyzed ADP- branes is enhanced by guanosine 5'-[v-thioltriphosphate ribosylation of the a subunit of the trimeric GTP-binding (GTP[yS]), a nonhydrolyzable analogue of GTP, and by a protein G, (G,.) (19). ARF is an abundant cytosolic protein mixture of aluminum and fluoride ions (Al/F). Here we show that reversibly associates with Golgi membranes (20, 21). that the ADP-ribosylation factor (ARF) is required for the ARF has been shown to be present on Golgi coated vesicles binding of (-COP. Thus, 13-COP contained in a coatomer generated in the presence of GTP[yS], but it is not a com- fraction that has been resolved from ARF does not bind to Golgi ponent of the cytosolic coatomer (22).
    [Show full text]
  • Mechanisms of Synaptic Plasticity Mediated by Clathrin Adaptor-Protein Complexes 1 and 2 in Mice
    Mechanisms of synaptic plasticity mediated by Clathrin Adaptor-protein complexes 1 and 2 in mice Dissertation for the award of the degree “Doctor rerum naturalium” at the Georg-August-University Göttingen within the doctoral program “Molecular Biology of Cells” of the Georg-August University School of Science (GAUSS) Submitted by Ratnakar Mishra Born in Birpur, Bihar, India Göttingen, Germany 2019 1 Members of the Thesis Committee Prof. Dr. Peter Schu Institute for Cellular Biochemistry, (Supervisor and first referee) University Medical Center Göttingen, Germany Dr. Hans Dieter Schmitt Neurobiology, Max Planck Institute (Second referee) for Biophysical Chemistry, Göttingen, Germany Prof. Dr. med. Thomas A. Bayer Division of Molecular Psychiatry, University Medical Center, Göttingen, Germany Additional Members of the Examination Board Prof. Dr. Silvio O. Rizzoli Department of Neuro-and Sensory Physiology, University Medical Center Göttingen, Germany Dr. Roland Dosch Institute of Developmental Biochemistry, University Medical Center Göttingen, Germany Prof. Dr. med. Martin Oppermann Institute of Cellular and Molecular Immunology, University Medical Center, Göttingen, Germany Date of oral examination: 14th may 2019 2 Table of Contents List of abbreviations ................................................................................. 5 Abstract ................................................................................................... 7 Chapter 1: Introduction ............................................................................
    [Show full text]
  • Palmitoylation: Implications for Nitric Oxide Signaling
    Proc. Natl. Acad. Sci. USA Vol. 93, pp. 6448-6453, June 1996 Cell Biology Targeting of nitric oxide synthase to endothelial cell caveolae via palmitoylation: Implications for nitric oxide signaling (endothelial nitric oxide synthase/signal transduction/vascular biology/N-myristoylation) GUILLERMO GARC1A-CARDENA*, PHIL OHt, JIANwEI LIu*, JAN E. SCHNITZERt, AND WILLIAM C. SESSA*t *Molecular Cardiobiology Program and Department of Pharmacology, Yale University School of Medicine, 295 Congress Avenue, New Haven, CT 06536; and tDepartment of Pathology, Harvard Medical School, Beth Israel Hospital, 330 Brookline Avenue, Boston, MA 02215 Communicated by Vincent T. Marchesi, Yale Univeristy, New Haven, CT, March 13, 1996 (received for review February 5, 1996) ABSTRACT The membrane association of endothelial insoluble membranes (TIM), suggesting that caveolae are nitric oxide synthase (eNOS) plays an important role in the signal processing centers (2-11). Additionally, caveolae have biosynthesis of nitric oxide (NO) in vascular endothelium. been implicated in other important cellular functions, includ- Previously, we have shown that in cultured endothelial cells ing endocytosis, potocytosis, and transcytosis (12, 13). and in intact blood vessels, eNOS is found primarily in the Endothelial nitric oxide synthase (eNOS) is a peripheral perinuclear region of the cells and in discrete regions of the membrane protein that metabolizes L-arginine to nitric oxide plasma membrane, suggesting trafficking of the protein from (NO). NO is a short-lived free radical gas involved in diverse the Golgi to specialized plasma membrane structures. Here, physiological and pathological processes. Endothelial-derived we show that eNOS is found in Triton X-100-insoluble mem- NO is an important paracrine mediator of vascular smooth branes prepared from cultured bovine aortic endothelial cells muscle tone and is an inhibitor of leukocyte adhesion and and colocalizes with caveolin, a coat protein of caveolae, in platelet aggregation (14, 15).
    [Show full text]
  • ADP-Ribosylation Factor and Coatomer Couple Fusion to Vesicle Budding Zvulun Elazar,* Lelio Orci,T Joachim Ostermann,* Myl~Nc Amherdt,* Gary Tanigawa,* and James E
    ADP-Ribosylation Factor and Coatomer Couple Fusion to Vesicle Budding Zvulun Elazar,* Lelio Orci,t Joachim Ostermann,* Myl~nc Amherdt,* Gary Tanigawa,* and James E. Rothman* * Program in Cellular Biochemistry and Biophysics, Memorial Sloan Kettering Cancer Center, New York 10021; and ¢Institute of Histology and Embryology, University of Geneva Medical School, 1211 Geneva 4, Switzerland Abstract. The coat proteins required for budding pair directly without an intervening vesicle. Coupling COP-coated vesicles from Golgi membranes, coatomer may therefore result from the sequestration of fuso- and ADP-ribosylation factor (ARF) protein, are shown genic membrane proteins into assembling coated vesi- to be required to reconstitute the orderly process of cles that are only exposed when the coat is removed transport between Golgi cisternae in which fusion of after budding is complete. This mechanism of cou- transport vesicles begins only after budding ends. pling explains the phenomenon of "retrograde transport" When either coat protein is omitted, fusion is uncou- triggered by uncouplers such as the drug brefeldin A. pled from budding-donor and acceptor compartments ow is membrane fusion coupled to vesicle budding? Orci et al., 1989). Purification of COP-coated vesicles (Mal- A transport vesicle must fuse with its target only hotra et al., 1989; Serafini et al., 1991a,b) revealed that their H after its budding from the parental membrane is coats consist of a small GTP-binding protein (ADP-ribosyla- completed. Otherwise, the various membrane-bound com- tion factor, ARF) ~ and a complex of seven distinct proteins partments connected by vesicle shuttles would fuse and the termed coatomer (Waters et al., 1992a; Stenbeck et al., topological organization of the endomembrane system in 1993), whose subunits are or, B, B', 3', ~, e, and ~'-COPs.
    [Show full text]
  • Endothelial Plasmalemma Vesicle–Associated Protein Regulates the Homeostasis of Splenic Immature B Cells and B-1 B Cells
    Endothelial Plasmalemma Vesicle−Associated Protein Regulates the Homeostasis of Splenic Immature B Cells and B-1 B Cells This information is current as Raul Elgueta, Dan Tse, Sophie J. Deharvengt, Marcus R. of September 26, 2021. Luciano, Catherine Carriere, Randolph J. Noelle and Radu V. Stan J Immunol 2016; 197:3970-3981; Prepublished online 14 October 2016; doi: 10.4049/jimmunol.1501859 Downloaded from http://www.jimmunol.org/content/197/10/3970 Supplementary http://www.jimmunol.org/content/suppl/2016/10/13/jimmunol.150185 Material 9.DCSupplemental http://www.jimmunol.org/ References This article cites 64 articles, 25 of which you can access for free at: http://www.jimmunol.org/content/197/10/3970.full#ref-list-1 Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision by guest on September 26, 2021 • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2016 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Endothelial Plasmalemma Vesicle–Associated Protein Regulates the Homeostasis of Splenic Immature B Cells and B-1 B Cells Raul Elgueta,*,† Dan Tse,‡,1 Sophie J.
    [Show full text]
  • Dynamin Action at the Final Stage of Clathrin- Mediated Endocytosis
    Dynamin Action at the Final Stage of Clathrin- Mediated Endocytosis Ning Fang ( [email protected] ) Georgia State University https://orcid.org/0000-0003-4710-0984 Xiaodong Cheng Georgia State University Kuangcai Chen Georgia State University https://orcid.org/0000-0002-9321-9225 Bin Dong Georgia State University https://orcid.org/0000-0002-3196-0712 Meek Yang Georgia State University Seth Filbrun Georgia State University Yong Myoung Georgia State University Teng-Xiang Huang Georgia State University Yan Gu The Bristol-Myers Squibb Company Gufeng Wang Georgia State University Article Keywords: Dynamin, clathrin-mediated endocytosis, scission, vesicle ssion Posted Date: February 5th, 2021 DOI: https://doi.org/10.21203/rs.3.rs-134570/v1 License: This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Page 1/25 Abstract Dynamin plays an important role in clathrin-mediated endocytosis by cutting the neck of nascent vesicles from the cell membrane. Gold nanorods were used as imaging probes to observe dynamin action on cargo vesicles during live endocytosis events. Invariant is that at the peak of dynamin accumulation, the cargo-containing vesicle always gives abrupt, right-handed rotations that nishes in a short time (~ 0.28 s). The large and quick twist, herein named the super twist, is the result of the coordinated dynamin helix action upon GTP hydrolysis. After the super twist, the rotational freedom of the vesicle drastically increases, accompanied with simultaneous or delayed translational movement, indicating that it detaches from the cell membrane. These observations suggest that dynamin-mediated scission at the nal stage involves a large torque generated by coordinated actions of multiple dynamins in the helix, which is the main driving force for scission.
    [Show full text]
  • The Structure of the COPI Coat Determined Within the Cell
    RESEARCH ADVANCE The structure of the COPI coat determined within the cell Yury S Bykov1,2, Miroslava Schaffer3, Svetlana O Dodonova1†, Sahradha Albert3, Ju¨ rgen M Plitzko3, Wolfgang Baumeister3*, Benjamin D Engel3*, John AG Briggs1,2* 1Structural and Computational Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany; 2Structural Studies Division, MRC Laboratory of Molecular Biology, Cambridge, United Kingdom; 3Department of Molecular Structural Biology, Max Planck Institute of Biochemistry, Martinsried, Germany Abstract COPI-coated vesicles mediate trafficking within the Golgi apparatus and from the Golgi to the endoplasmic reticulum. The structures of membrane protein coats, including COPI, have been extensively studied with in vitro reconstitution systems using purified components. Previously we have determined a complete structural model of the in vitro reconstituted COPI coat (Dodonova et al., 2017). Here, we applied cryo-focused ion beam milling, cryo-electron tomography and subtomogram averaging to determine the native structure of the COPI coat within vitrified Chlamydomonas reinhardtii cells. The native algal structure resembles the in vitro mammalian structure, but additionally reveals cargo bound beneath b’–COP. We find that all coat components *For correspondence: disassemble simultaneously and relatively rapidly after budding. Structural analysis in situ, [email protected] (WB); maintaining Golgi topology, shows that vesicles change their size, membrane thickness, and cargo [email protected] (BDE); content as they progress from cis to trans, but the structure of the coat machinery remains [email protected] constant. (JAGB) DOI: https://doi.org/10.7554/eLife.32493.001 Present address: †Department of Molecular Biology, Max Planck Institute for Biophysical Chemistry, Go¨ ttingen, Germany Introduction Vesicular transport between cellular compartments is a fundamental mechanism of eukaryotic cells.
    [Show full text]