Home LiteratureArticle Details
PMID: 11950941 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Bilayered clathrin coats on endosomal vacuoles are involved in protein sorting toward lysosomes.

Molecular biology of the cell ·Vol. 13 ·No. 4 ·2002-04-00 ·Pages 1313-28

Sachse M, Urbé S, Oorschot V, Strous GJ, Klumperman J

Abstract

In many cells endosomal vacuoles show clathrin coats of which the function is unknown. Herein, we show that this coat is predominantly present on early endosomes and has a characteristic bilayered appearance in the electron microscope. By immunoelectron microscopy we show that the coat contains clathrin heavy as well as light chain, but lacks the adaptor complexes AP1, AP2, and AP3, by which it differs from clathrin coats on endocytic vesicles and recycling endosomes. The coat is insensitive to short incubations with brefeldin A, but disappears in the presence of the phosphatidylinositol 3-kinase inhibitor wortmannin. No association of endosomal coated areas with tracks of tubulin or actin was found. By quantitative immunoelectron microscopy, we found that the lysosomal-targeted receptors for growth hormone (GHR) and epidermal growth factor are concentrated in the coated membrane areas, whereas the recycling transferrin receptor is not. In addition, we found that the proteasomal inhibitor MG 132 induces a redistribution of a truncated GHR (GHR-369) toward recycling vesicles, which coincided with a redistribution of endosomal vacuole-associated GHR-369 to the noncoated areas of the limiting membrane. Together, these data suggest a role for the bilayered clathrin coat on vacuolar endosomes in targeting of proteins to lysosomes.

MeSH Terms
Androstadienes/pharmacology Animals Brefeldin A/pharmacology CHO Cells Cell Membrane/metabolism Clathrin/chemistry,metabolism Cricetinae Cytoskeleton/metabolism Endocytosis Endosomes/metabolism Enzyme Inhibitors/pharmacology HeLa Cells Humans Kinetics Ligands Lipid Bilayers/metabolism Lysosomes/metabolism Membrane Proteins/metabolism Models, Biological Protein Binding Qa-SNARE Proteins Time Factors Transferrin/metabolism Wortmannin
Chemicals
Androstadienes Clathrin Enzyme Inhibitors Ligands Lipid Bilayers Membrane Proteins Qa-SNARE Proteins Transferrin Brefeldin A Wortmannin
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Sachse Martin
Department of Cell Biology, University Medical Center Utrecht and Institute of Biomembranes, 3584 CX Utrecht, The Netherlands.
Urbé Sylvie
Oorschot Viola
Strous Ger J
Klumperman Judith
References (81)
81 references, click to expand
  1. Histochemical detection of binding sites for human growth hormone using biotinylated ligand.
    J Histochem Cytochem. 1994 Jan;42(1):103-7 PMID: 8263321
  2. In migrating fibroblasts, recycling receptors are concentrated in narrow tubules in the pericentriolar area, and then routed to the plasma membrane of the leading lamella.
    J Cell Biol. 1994 Jun;125(6):1265-74 PMID: 7515888
  3. Delivery to lysosomes in the human carcinoma cell line HEp-2 involves an actin filament-facilitated fusion between mature endosomes and preexisting lysosomes.
    Eur J Cell Biol. 1995 Apr;66(4):309-23 PMID: 7544728
  4. Tyrosine phosphorylation of the c-cbl proto-oncogene protein product and association with epidermal growth factor (EGF) receptor upon EGF stimulation.
    J Biol Chem. 1995 Sep 1;270(35):20242-5 PMID: 7657591
  5. Syntaxin 7 and VAMP-7 are soluble N-ethylmaleimide-sensitive factor attachment protein receptors required for late endosome-lysosome and homotypic lysosome fusion in alveolar macrophages.
    Mol Biol Cell. 2000 Jul;11(7):2327-33 PMID: 10888671
  6. VPS27 controls vacuolar and endocytic traffic through a prevacuolar compartment in Saccharomyces cerevisiae.
    J Cell Biol. 1995 Nov;131(3):603-17 PMID: 7593183
  7. Association of the AP-3 adaptor complex with clathrin.
    Science. 1998 Apr 17;280(5362):431-4 PMID: 9545220
  8. In polarized MDCK cells basolateral vesicles arise from clathrin-gamma-adaptin-coated domains on endosomal tubules.
    J Cell Biol. 1998 May 4;141(3):611-23 PMID: 9566963
  9. Di-leucine-mediated internalization of ligand by a truncated growth hormone receptor is independent of the ubiquitin conjugation system.
    J Biol Chem. 1998 Jun 26;273(26):16426-33 PMID: 9632708
  10. ADP-Ribosylation factor 1 (ARF1) regulates recruitment of the AP-3 adaptor complex to membranes.
    J Cell Biol. 1998 Jul 27;142(2):391-402 PMID: 9679139
  11. Molecular characterization of the signal responsible for the targeting of the interleukin 2 receptor beta chain toward intracellular degradation.
    J Biol Chem. 1998 Nov 6;273(45):29424-9 PMID: 9792646
  12. Syntaxin 13 mediates cycling of plasma membrane proteins via tubulovesicular recycling endosomes.
    J Cell Biol. 1998 Nov 16;143(4):957-71 PMID: 9817754
  13. Direct pathway from early/recycling endosomes to the Golgi apparatus revealed through the study of shiga toxin B-fragment transport.
    J Cell Biol. 1998 Nov 16;143(4):973-90 PMID: 9817755
  14. c-Cbl/Sli-1 regulates endocytic sorting and ubiquitination of the epidermal growth factor receptor.
    Genes Dev. 1998 Dec 1;12(23):3663-74 PMID: 9851973
  15. Fab1p PtdIns(3)P 5-kinase function essential for protein sorting in the multivesicular body.
    Cell. 1998 Dec 11;95(6):847-58 PMID: 9865702
  16. Identification of a novel ubiquitin conjugation motif, required for ligand-induced internalization of the growth hormone receptor.
    EMBO J. 1999 Jan 4;18(1):28-36 PMID: 9878047
  17. Clathrin, adaptors and eps15 in endosomes containing activated epidermal growth factor receptors.
    J Cell Sci. 1999 Feb;112 ( Pt 3):317-27 PMID: 9885285
  18. A novel class of clathrin-coated vesicles budding from endosomes.
    J Cell Biol. 1996 Jan;132(1-2):21-33 PMID: 8567724
  19. The ubiquitin conjugation system is required for ligand-induced endocytosis and degradation of the growth hormone receptor.
    EMBO J. 1996 Aug 1;15(15):3806-12 PMID: 8670885
  20. Improving structural integrity of cryosections for immunogold labeling.
    Histochem Cell Biol. 1996 Jul;106(1):41-58 PMID: 8858366
  21. Wortmannin alters the transferrin receptor endocytic pathway in vivo and in vitro.
    Mol Biol Cell. 1996 Mar;7(3):355-67 PMID: 8868465
  22. Lysosomal targeting of epidermal growth factor receptors via a kinase-dependent pathway is mediated by the receptor carboxyl-terminal residues 1022-1123.
    J Biol Chem. 1996 Nov 29;271(48):30340-6 PMID: 8939994
  23. AP-2-containing clathrin coats assemble on mature lysosomes.
    J Cell Biol. 1996 Dec;135(6 Pt 2):1801-14 PMID: 8991092
  24. Clathrin-coated vesicle formation and protein sorting: an integrated process.
    Annu Rev Biochem. 1997;66:511-48 PMID: 9242916
  25. Hrs, a tyrosine kinase substrate with a conserved double zinc finger domain, is localized to the cytoplasmic surface of early endosomes.
    J Biol Chem. 1997 Aug 15;272(33):20538-44 PMID: 9252367
  26. Linkage of the ubiquitin-conjugating system and the endocytic pathway in ligand-induced internalization of the growth hormone receptor.
    EMBO J. 1997 Aug 15;16(16):4851-8 PMID: 9305627
  27. Major histocompatibility complex class II compartments in human and mouse B lymphoblasts represent conventional endocytic compartments.
    J Cell Biol. 1997 Nov 3;139(3):639-49 PMID: 9348281
  28. Syntaxin 7, a novel syntaxin member associated with the early endosomal compartment.
    J Biol Chem. 1998 Jan 2;273(1):375-80 PMID: 9417091
  29. Localization of phosphatidylinositol 3-phosphate in yeast and mammalian cells.
    EMBO J. 2000 Sep 1;19(17):4577-88 PMID: 10970851
  30. Syntaxin 7 is localized to late endosome compartments, associates with Vamp 8, and Is required for late endosome-lysosome fusion.
    Mol Biol Cell. 2000 Sep;11(9):3137-53 PMID: 10982406
  31. Endosomal localization and receptor dynamics determine tyrosine phosphorylation of hepatocyte growth factor-regulated tyrosine kinase substrate.
    Mol Cell Biol. 2000 Oct;20(20):7685-92 PMID: 11003664
  32. A deubiquitinating enzyme UBPY interacts with the Src homology 3 domain of Hrs-binding protein via a novel binding motif PX(V/I)(D/N)RXXKP.
    J Biol Chem. 2000 Dec 1;275(48):37481-7 PMID: 10982817
  33. A SNARE complex mediating fusion of late endosomes defines conserved properties of SNARE structure and function.
    EMBO J. 2000 Dec 1;19(23):6453-64 PMID: 11101518
  34. Immunolocalization of cytoplasmic dynein and dynactin subunits in cultured macrophages: enrichment on early endocytic organelles.
    J Cell Sci. 2001 Jan;114(Pt 1):229-240 PMID: 11112706
  35. A novel clathrin homolog that co-distributes with cytoskeletal components functions in the trans-Golgi network.
    EMBO J. 2001 Jan 15;20(1-2):272-84 PMID: 11226177
  36. Distinct protein sorting and localization to premelanosomes, melanosomes, and lysosomes in pigmented melanocytic cells.
    J Cell Biol. 2001 Feb 19;152(4):809-24 PMID: 11266471
  37. c-Cbl ubiquitinates the EGF receptor at the plasma membrane and remains receptor associated throughout the endocytic route.
    J Cell Sci. 2001 Jun;114(Pt 11):2167-78 PMID: 11493652
  38. The ubiquitin-proteasome pathway regulates lysosomal degradation of the growth hormone receptor and its ligand.
    Biochem Soc Trans. 2001 Aug;29(Pt 4):488-93 PMID: 11498015
  39. Ubiquitin-dependent sorting into the multivesicular body pathway requires the function of a conserved endosomal protein sorting complex, ESCRT-I.
    Cell. 2001 Jul 27;106(2):145-55 PMID: 11511343
  40. Proteasome inhibitors block a late step in lysosomal transport of selected membrane but not soluble proteins.
    Mol Biol Cell. 2001 Aug;12(8):2556-66 PMID: 11514635
  41. Hrs recruits clathrin to early endosomes.
    EMBO J. 2001 Sep 3;20(17):5008-21 PMID: 11532964
  42. Ubiquitin sorts proteins into the intralumenal degradative compartment of the late-endosome/vacuole.
    Traffic. 2001 Sep;2(9):622-30 PMID: 11555416
  43. Sorting of proteins into multivesicular bodies: ubiquitin-dependent and -independent targeting.
    EMBO J. 2001 Sep 17;20(18):5176-86 PMID: 11566881
  44. Regulation of receptor fate by ubiquitination of activated beta 2-adrenergic receptor and beta-arrestin.
    Science. 2001 Nov 9;294(5545):1307-13 PMID: 11588219
  45. The ubiquitin-dependent endocytosis motif is required for efficient incorporation of growth hormone receptor in clathrin-coated pits, but not clathrin-coated lattices.
    J Cell Sci. 2001 Nov;114(Pt 21):3943-52 PMID: 11719561
  46. Agonist-promoted ubiquitination of the G protein-coupled receptor CXCR4 mediates lysosomal sorting.
    J Biol Chem. 2001 Dec 7;276(49):45509-12 PMID: 11641392
  47. Dynamin-dependent transferrin receptor recycling by endosome-derived clathrin-coated vesicles.
    Mol Biol Cell. 2002 Jan;13(1):169-82 PMID: 11809831
  48. Cytochemical studies of lysosomes, golgi apparatus and endoplasmic reticulum in secretion and protein uptake by adrenal medulla cells of the rat.
    J Histochem Cytochem. 1968 May;16(5):320-36 PMID: 5656143
  49. Biosynthesis of the human transferrin receptor in cultured cells.
    J Biol Chem. 1981 Dec 25;256(24):12888-92 PMID: 6273413
  50. Intracellular site of asialoglycoprotein receptor-ligand uncoupling: double-label immunoelectron microscopy during receptor-mediated endocytosis.
    Cell. 1983 Jan;32(1):277-87 PMID: 6130851
  51. Receptor-mediated endocytosis of epidermal growth factor by rat hepatocytes: receptor pathway.
    J Cell Biol. 1986 Jan;102(1):24-36 PMID: 3001101
  52. Growth hormone receptors in cultured adipocytes: a model to study receptor regulation.
    Mol Cell Endocrinol. 1986 Sep;47(1-2):81-90 PMID: 3017789
  53. Sorting of endocytosed transferrin and asialoglycoprotein occurs immediately after internalization in HepG2 cells.
    J Cell Biol. 1987 May;104(5):1261-8 PMID: 3032986
  54. Translocation and clustering of endosomes and lysosomes depends on microtubules.
    J Cell Biol. 1987 Sep;105(3):1253-65 PMID: 3308906
  55. A Chinese hamster cell cycle mutant arrested at G2 phase has a temperature-sensitive ubiquitin-activating enzyme, E1.
    J Biol Chem. 1988 Oct 25;263(30):15726-31 PMID: 3049611
  56. A quantitative model of traffic between plasma membrane and secondary lysosomes: evaluation of inflow, lateral diffusion, and degradation.
    J Cell Biol. 1988 Dec;107(6 Pt 1):2109-15 PMID: 2848849
  57. Iterative fractionation of recycling receptors from lysosomally destined ligands in an early sorting endosome.
    J Cell Biol. 1989 Dec;109(6 Pt 2):3303-14 PMID: 2600137
  58. Role of the human transferrin receptor cytoplasmic domain in endocytosis: localization of a specific signal sequence for internalization.
    J Cell Biol. 1990 Feb;110(2):283-94 PMID: 2298808
  59. Kinase activity controls the sorting of the epidermal growth factor receptor within the multivesicular body.
    Cell. 1990 May 18;61(4):623-34 PMID: 2344614
  60. Immuno-localization of the insulin regulatable glucose transporter in brown adipose tissue of the rat.
    J Cell Biol. 1991 Apr;113(1):123-35 PMID: 2007617
  61. Recruitment of coat proteins onto Golgi membranes in intact and permeabilized cells: effects of brefeldin A and G protein activators.
    Cell. 1992 Apr 3;69(1):129-38 PMID: 1555237
  62. Differences in the endosomal distributions of the two mannose 6-phosphate receptors.
    J Cell Biol. 1993 Jun;121(5):997-1010 PMID: 8099077
  63. Multivesicular body morphogenesis requires phosphatidyl-inositol 3-kinase activity.
    Curr Biol. 1999 Jan 14;9(1):55-8 PMID: 9889123
  64. A leucine-based determinant in the epidermal growth factor receptor juxtamembrane domain is required for the efficient transport of ligand-receptor complexes to lysosomes.
    J Biol Chem. 1999 Jan 29;274(5):3141-50 PMID: 9915853
  65. A tubular endosomal fraction from rat liver: biochemical evidence of receptor sorting by default.
    Proc Natl Acad Sci U S A. 1999 Aug 31;96(18):10146-51 PMID: 10468577
  66. The tyrosine kinase negative regulator c-Cbl as a RING-type, E2-dependent ubiquitin-protein ligase.
    Science. 1999 Oct 8;286(5438):309-12 PMID: 10514377
  67. Differential roles of syntaxin 7 and syntaxin 8 in endosomal trafficking.
    Mol Biol Cell. 1999 Nov;10(11):3891-908 PMID: 10564279
  68. Phosphatidylinositol-3-OH kinases are Rab5 effectors.
    Nat Cell Biol. 1999 Aug;1(4):249-52 PMID: 10559924
  69. Synaptic vesicles form by budding from tubular extensions of sorting endosomes in PC12 cells.
    Mol Biol Cell. 1999 Dec;10(12):4163-76 PMID: 10588650
  70. Clathrin-coated lattices and buds on MHC class II compartments do not selectively recruit mature MHC-II.
    J Cell Sci. 2000 Jan;113 Pt 2:303-13 PMID: 10633081
  71. Ubiquitin ligase activity and tyrosine phosphorylation underlie suppression of growth factor signaling by c-Cbl/Sli-1.
    Mol Cell. 1999 Dec;4(6):1029-40 PMID: 10635327
  72. Endocytosis and degradation of the growth hormone receptor are proteasome-dependent.
    J Biol Chem. 2000 Jan 21;275(3):1575-80 PMID: 10636847
  73. A hrs binding protein having a Src homology 3 domain is involved in intracellular degradation of growth factors and their receptors.
    Genes Cells. 2000 Jan;5(1):57-69 PMID: 10651905
  74. Selective membrane recruitment of EEA1 suggests a role in directional transport of clathrin-coated vesicles to early endosomes.
    J Biol Chem. 2000 Feb 11;275(6):3745-8 PMID: 10660521
  75. Actin-dependent propulsion of endosomes and lysosomes by recruitment of N-WASP.
    J Cell Biol. 2000 Feb 7;148(3):519-30 PMID: 10662777
  76. Syntaxin 7 mediates endocytic trafficking to late endosomes.
    J Biol Chem. 2000 Mar 3;275(9):6523-9 PMID: 10692457
  77. Polyubiquitination of the epidermal growth factor receptor occurs at the plasma membrane upon ligand-induced activation.
    J Biol Chem. 2000 May 5;275(18):13940-7 PMID: 10788520
  78. Sorting in the endosomal system in yeast and animal cells.
    Curr Opin Cell Biol. 2000 Aug;12(4):457-66 PMID: 10873832
  79. Role of auxilin in uncoating clathrin-coated vesicles.
    Nature. 1995 Dec 7;378(6557):632-5 PMID: 8524399
  80. Sorting of membrane components from endosomes and subsequent recycling to the cell surface occurs by a bulk flow process.
    J Cell Biol. 1993 Jun;121(6):1257-69 PMID: 8509447
  81. Multivesicular bodies in HEp-2 cells are maturing endosomes.
    Eur J Cell Biol. 1993 Aug;61(2):208-24 PMID: 8223712
Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.01-10-0525
Published
2002-04-00
Pages
1313-28
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC102271
Subset
IM
Grants
NHLBI NIH HHS · HL-59150 · United States
NINDS NIH HHS · NS-37525 · United States
External Links
PubMed source DOI: 10.1091/mbc.01-10-0525 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2