Home LiteratureArticle Details
PMID: 9456319 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Fusion of lysosomes with late endosomes produces a hybrid organelle of intermediate density and is NSF dependent.

The Journal of cell biology ·Vol. 140 ·No. 3 ·1998-02-09 ·Pages 591-601

Mullock BM, Bright NA, Fearon CW, Gray SR, Luzio JP

Abstract

Using a cell-free content mixing assay containing rat liver endosomes and lysosomes in the presence of pig brain cytosol, we demonstrated that after incubation at 37 degrees C, late endosome-lysosome hybrid organelles were formed, which could be isolated by density gradient centrifugation. ImmunoEM showed that the hybrids contained both an endocytosed marker and a lysosomal enzyme. Formation of the hybrid organelles appeared not to require vesicular transport between late endosomes and lysosomes but occurred as a result of direct fusion. Hybrid organelles with similar properties were isolated directly from rat liver homogenates and thus were not an artifact of cell-free incubations. Direct fusion between late endosomes and lysosomes was an N-ethylmaleimide-sensitive factor-dependent event and was inhibited by GDP-dissociation inhibitor, indicating a requirement for a rab protein. We suggest that in cells, delivery of endocytosed ligands to an organelle where proteolytic digestion occurs is mediated by direct fusion of late endosomes with lysosomes. The consequences of this fusion to the maintenance and function of lysosomes are discussed.

MeSH Terms
Adenosine Triphosphate/metabolism Androstadienes/pharmacology Animals Carrier Proteins/pharmacology Cell-Free System Centrifugation, Density Gradient Chromones/pharmacology Cytosol/physiology Endocytosis Endosomes/physiology,ultrastructure Enzyme Inhibitors/pharmacology Ethylmaleimide/pharmacology GTP-Binding Proteins/physiology Guanine Nucleotide Dissociation Inhibitors Guanosine Triphosphate/metabolism Liver/ultrastructure Lysosomes/physiology,ultrastructure Membrane Fusion Membrane Proteins/pharmacology Morpholines/pharmacology N-Ethylmaleimide-Sensitive Proteins Phosphoinositide-3 Kinase Inhibitors Rats Recombinant Proteins/pharmacology Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins Swine Vesicular Transport Proteins Wortmannin
Chemicals
Androstadienes Carrier Proteins Chromones Enzyme Inhibitors GDP dissociation inhibitor 1 Guanine Nucleotide Dissociation Inhibitors Membrane Proteins Morpholines Phosphoinositide-3 Kinase Inhibitors Recombinant Proteins Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins Vesicular Transport Proteins 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one Guanosine Triphosphate Adenosine Triphosphate GTP-Binding Proteins N-Ethylmaleimide-Sensitive Proteins Nsf protein, rat Ethylmaleimide Wortmannin
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Mullock B M
Department of Clinical Biochemistry, University of Cambridge, Addenbrooke's Hospital, Cambridge, CB2 2QR, United Kingdom.
Bright N A
Fearon C W
Gray S R
Luzio J P
References (51)
51 references, click to expand
  1. 3-Methyladenine inhibits transport from late endosomes to lysosomes in cultured rat and mouse fibroblasts.
    Eur J Cell Biol. 1994 Oct;65(1):14-25 PMID: 7889984
  2. Homotypic lysosome fusion in macrophages: analysis using an in vitro assay.
    J Cell Biol. 1997 Nov 3;139(3):665-73 PMID: 9348283
  3. Rab GDP dissociation inhibitor: putting rab GTPases in the right place.
    J Biol Chem. 1995 Jul 21;270(29):17057-9 PMID: 7615494
  4. Transport from late endosomes to lysosomes, but not sorting of integral membrane proteins in endosomes, depends on the vacuolar proton pump.
    J Cell Biol. 1995 Aug;130(4):821-34 PMID: 7642700
  5. 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
  6. Membrane transport in the endocytic pathway.
    Curr Opin Cell Biol. 1995 Aug;7(4):552-63 PMID: 7495576
  7. The formation of Golgi stacks from vesiculated Golgi membranes requires two distinct fusion events.
    Cell. 1995 Sep 22;82(6):895-904 PMID: 7553850
  8. An NSF-like ATPase, p97, and NSF mediate cisternal regrowth from mitotic Golgi fragments.
    Cell. 1995 Sep 22;82(6):905-14 PMID: 7553851
  9. Evidence for phosphatidylinositol 3-kinase as a regulator of endocytosis via activation of Rab5.
    Proc Natl Acad Sci U S A. 1995 Oct 24;92(22):10207-11 PMID: 7479754
  10. Dense core lysosomes can fuse with late endosomes and are re-formed from the resultant hybrid organelles.
    J Cell Sci. 1997 Sep;110 ( Pt 17):2027-40 PMID: 9378754
  11. Use of Rab-GDP dissociation inhibitor for solubilization and delivery of Rab proteins to biological membranes in streptolysin O-permeabilized cells.
    Methods Enzymol. 1995;257:243-53 PMID: 8583927
  12. Cytoplasmic coat proteins involved in endosome function.
    Cell. 1995 Dec 1;83(5):703-13 PMID: 8521487
  13. Rab 7: an important regulator of late endocytic membrane traffic.
    J Cell Biol. 1995 Dec;131(6 Pt 1):1435-52 PMID: 8522602
  14. A novel class of clathrin-coated vesicles budding from endosomes.
    J Cell Biol. 1996 Jan;132(1-2):21-33 PMID: 8567724
  15. The rab7 GTPase resides on a vesicular compartment connected to lysosomes.
    J Cell Sci. 1995 Nov;108 ( Pt 11):3349-58 PMID: 8586647
  16. Multivesicular endosomes containing internalized EGF-EGF receptor complexes mature and then fuse directly with lysosomes.
    J Cell Biol. 1996 Mar;132(6):1011-23 PMID: 8601581
  17. Thioredoxin is required for vacuole inheritance in Saccharomyces cerevisiae.
    J Cell Biol. 1996 Mar;132(5):787-94 PMID: 8603912
  18. An endosomal beta COP is involved in the pH-dependent formation of transport vesicles destined for late endosomes.
    J Cell Biol. 1996 Apr;133(1):29-41 PMID: 8601610
  19. Association of the fusion protein NSF with clathrin-coated vesicle membranes.
    EMBO J. 1996 Feb 15;15(4):745-52 PMID: 8631296
  20. The effect of wortmannin on the localisation of lysosomal type I integral membrane glycoproteins suggests a role for phosphoinositide 3-kinase activity in regulating membrane traffic late in the endocytic pathway.
    J Cell Sci. 1996 Apr;109 ( Pt 4):749-62 PMID: 8718666
  21. Actin filaments facilitate two steps of endocytosis.
    J Cell Sci. 1996 Feb;109 ( Pt 2):457-65 PMID: 8838669
  22. Improving structural integrity of cryosections for immunogold labeling.
    Histochem Cell Biol. 1996 Jul;106(1):41-58 PMID: 8858366
  23. The biogenesis of lysosomes: is it a kiss and run, continuous fusion and fission process?
    Bioessays. 1996 Nov;18(11):895-903 PMID: 8939067
  24. Endocytosis and molecular sorting.
    Annu Rev Cell Dev Biol. 1996;12:575-625 PMID: 8970738
  25. AP-2-containing clathrin coats assemble on mature lysosomes.
    J Cell Biol. 1996 Dec;135(6 Pt 2):1801-14 PMID: 8991092
  26. A study of positive staining of ultrathin frozen sections.
    J Ultrastruct Res. 1978 Jun;63(3):287-307 PMID: 79660
  27. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase.
    Gene. 1988 Jul 15;67(1):31-40 PMID: 3047011
  28. Identification of a novel, N-ethylmaleimide-sensitive cytosolic factor required for vesicular transport from endosomes to the trans-Golgi network in vitro.
    J Cell Biol. 1991 Mar;112(5):823-31 PMID: 1999460
  29. 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
  30. Cell-free fusion of endocytic vesicles is regulated by phosphorylation.
    J Cell Biol. 1992 Jan;116(2):331-8 PMID: 1309811
  31. A multisubunit particle implicated in membrane fusion.
    J Cell Biol. 1992 May;117(3):531-8 PMID: 1315316
  32. Expression and purification of recombinant N-ethylmaleimide-sensitive fusion protein from Escherichia coli.
    Methods Enzymol. 1992;219:309-18 PMID: 1488003
  33. Rab9 functions in transport between late endosomes and the trans Golgi network.
    EMBO J. 1993 Feb;12(2):677-82 PMID: 8440258
  34. Small GTP-binding proteins on rat liver lysosomal membranes.
    Cell Struct Funct. 1992 Dec;17(6):363-9 PMID: 1295693
  35. SNAP family of NSF attachment proteins includes a brain-specific isoform.
    Nature. 1993 Mar 25;362(6418):353-5 PMID: 8455721
  36. Rab GDP dissociation inhibitor as a general regulator for the membrane association of rab proteins.
    J Biol Chem. 1993 Aug 25;268(24):18143-50 PMID: 8349690
  37. Targeting and mistargeting of plasma membrane adaptors in vitro.
    J Cell Biol. 1993 Dec;123(5):1093-105 PMID: 8245120
  38. Cytoplasmic dynein-dependent vesicular transport from early to late endosomes.
    J Cell Biol. 1993 Dec;123(6 Pt 1):1373-87 PMID: 8253838
  39. Rab GTPases in vesicular transport.
    Curr Opin Cell Biol. 1993 Aug;5(4):613-20 PMID: 8257602
  40. Lysosomes can fuse with a late endosomal compartment in a cell-free system from rat liver.
    J Cell Biol. 1994 Sep;126(5):1173-82 PMID: 7520447
  41. Mechanisms of intracellular protein transport.
    Nature. 1994 Nov 3;372(6501):55-63 PMID: 7969419
  42. Rab GTPases: master regulators of membrane trafficking.
    Curr Opin Cell Biol. 1994 Aug;6(4):522-6 PMID: 7986528
  43. Multiple N-ethylmaleimide-sensitive components are required for endosomal vesicle fusion.
    Mol Biol Cell. 1994 Jul;5(7):773-83 PMID: 7812046
  44. The autophagic and endocytic pathways converge at the nascent autophagic vacuoles.
    J Cell Biol. 1997 Jan 13;136(1):61-70 PMID: 9008703
  45. The small GTP binding protein rab7 is essential for cellular vacuolation induced by Helicobacter pylori cytotoxin.
    EMBO J. 1997 Jan 2;16(1):15-24 PMID: 9009263
  46. Isolation and characterization of early endosomes, late endosomes and terminal lysosomes: their role in protein degradation.
    J Cell Sci. 1996 Dec;109 ( Pt 12):2905-14 PMID: 9013338
  47. Transport through the yeast endocytic pathway occurs through morphologically distinct compartments and requires an active secretory pathway and Sec18p/N-ethylmaleimide-sensitive fusion protein.
    Mol Biol Cell. 1997 Jan;8(1):13-31 PMID: 9017592
  48. The phosphatidylinositol 3-kinase inhibitors wortmannin and LY294002 inhibit autophagy in isolated rat hepatocytes.
    Eur J Biochem. 1997 Jan 15;243(1-2):240-6 PMID: 9030745
  49. Lysosomes behave as Ca2+-regulated exocytic vesicles in fibroblasts and epithelial cells.
    J Cell Biol. 1997 Apr 7;137(1):93-104 PMID: 9105039
  50. Reconstitution of phagosome-lysosome fusion in streptolysin O-permeabilized cells.
    J Biol Chem. 1997 Jun 27;272(26):16147-51 PMID: 9195911
  51. Physiological functions of endosomal proteolysis.
    Biochem J. 1995 Apr 15;307 ( Pt 2):313-26 PMID: 7733863
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.140.3.591
Published
1998-02-09
Pages
591-601
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2140175
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.140.3.591 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