J. Cell Biol., Volume 140, Number 3, February 9, 1998 591-601

Fusion of Lysosomes with Late Endosomes Produces a Hybrid Organelle of Intermediate Density and Is NSF Dependent

Barbara M. Mullock, Nicholas A. Bright, Clare W. Fearon, Sally R. Gray, and J. Luzio

Department of Clinical Biochemistry, University of Cambridge, Addenbrooke's Hospital, Cambridge, CB2 2QR, United Kingdom

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°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.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

• McNatt, M. W., McKittrick, I., West, M., Odorizzi, G. (2007). Direct Binding to Rsp5 Mediates Ubiquitin-independent Sorting of Sna3 via the Multivesicular Body Pathway. Mol. Biol. Cell 18: 697-706 [Abstract] [Full Text]  
• Odorizzi, G. (2006). The multiple personalities of Alix.. J. Cell Sci. 119: 3025-3032 [Abstract] [Full Text]  
• Razi, M., Futter, C. E. (2006). Distinct Roles for Tsg101 and Hrs in Multivesicular Body Formation and Inward Vesiculation. Mol. Biol. Cell 17: 3469-3483 [Abstract] [Full Text]  
• Tian, J.-H., Wu, Z.-X., Unzicker, M., Lu, L., Cai, Q., Li, C., Schirra, C., Matti, U., Stevens, D., Deng, C., Rettig, J., Sheng, Z.-H. (2005). The Role of Snapin in Neurosecretion: Snapin Knock-Out Mice Exhibit Impaired Calcium-Dependent Exocytosis of Large Dense-Core Vesicles in Chromaffin Cells. J. Neurosci. 25: 10546-10555 [Abstract] [Full Text]  
• Britton, C., Murray, L. (2004). Cathepsin L protease (CPL-1) is essential for yolk processing during embryogenesis in Caenorhabditis elegans. J. Cell Sci. 117: 5133-5143 [Abstract] [Full Text]  
• Treusch, S., Knuth, S., Slaugenhaupt, S. A., Goldin, E., Grant, B. D., Fares, H. (2004). Caenorhabditis elegans functional orthologue of human protein h-mucolipin-1 is required for lysosome biogenesis. Proc. Natl. Acad. Sci. USA 101: 4483-4488 [Abstract] [Full Text]  
• Vergne, I., Fratti, R. A., Hill, P. J., Chua, J., Belisle, J., Deretic, V. (2004). Mycobacterium tuberculosis Phagosome Maturation Arrest: Mycobacterial Phosphatidylinositol Analog Phosphatidylinositol Mannoside Stimulates Early Endosomal Fusion. Mol. Biol. Cell 15: 751-760 [Abstract] [Full Text]  
• Henry, R. M., Hoppe, A. D., Joshi, N., Swanson, J. A. (2004). The uniformity of phagosome maturation in macrophages. J. Cell Biol. 164: 185-194 [Abstract] [Full Text]  
• Murk, J. L. A. N., Humbel, B. M., Ziese, U., Griffith, J. M., Posthuma, G., Slot, J. W., Koster, A. J., Verkleij, A. J., Geuze, H. J., Kleijmeer, M. J. (2003). Endosomal compartmentalization in three dimensions: Implications for membrane fusion. Proc. Natl. Acad. Sci. USA 100: 13332-13337 [Abstract] [Full Text]  
• Chattopadhyay, S., Sun, P., Wang, P., Abonyo, B., Cross, N. L., Liu, L. (2003). Fusion of Lamellar Body with Plasma Membrane Is Driven by the Dual Action of Annexin II Tetramer and Arachidonic Acid. J. Biol. Chem. 278: 39675-39683 [Abstract] [Full Text]  
• Poupon, V., Stewart, A., Gray, S. R., Piper, R. C., Luzio, J. P. (2003). The Role of mVps18p in Clustering, Fusion, and Intracellular Localization of Late Endocytic Organelles. Mol. Biol. Cell 14: 4015-4027 [Abstract] [Full Text]  
• Duclos, S., Corsini, R., Desjardins, M. (2003). Remodeling of endosomes during lysosome biogenesis involves `kiss and run' fusion events regulated by rab5. J. Cell Sci. 116: 907-918 [Abstract] [Full Text]  
• Fujita, H., Yamanaka, M., Imamura, K., Tanaka, Y., Nara, A., Yoshimori, T., Yokota, S., Himeno, M. (2003). A dominant negative form of the AAA ATPase SKD1/VPS4 impairs membrane trafficking out of endosomal/lysosomal compartments: class E vps phenotype in mammalian cells. J. Cell Sci. 116: 401-414 [Abstract] [Full Text]  
• Croizet-Berger, K., Daumerie, C., Couvreur, M., Courtoy, P. J., van den Hove, M.-F. (2002). The endocytic catalysts, Rab5a and Rab7, are tandem regulators of thyroid hormone production. Proc. Natl. Acad. Sci. USA 99: 8277-8282 [Abstract] [Full Text]  
• Kuronita, T., Eskelinen, E.-L., Fujita, H., Saftig, P., Himeno, M., Tanaka, Y. (2002). A role for the lysosomal membrane protein LGP85 in the biogenesis and maintenance of endosomal and lysosomal morphology. J. Cell Sci. 115: 4117-4131 [Abstract] [Full Text]  
• Cordonnier, M.-N., Dauzonne, D., Louvard, D., Coudrier, E. (2001). Actin Filaments and Myosin I Alpha Cooperate with Microtubules for the Movement of Lysosomes. Mol. Biol. Cell 12: 4013-4029 [Abstract] [Full Text]  
• Webb, J. L., Harvey, M. W., Holden, D. W., Evans, T. J. (2001). Macrophage Nitric Oxide Synthase Associates with Cortical Actin but Is Not Recruited to Phagosomes. Infect. Immun. 69: 6391-6400 [Abstract] [Full Text]  
• Caplan, S., Hartnell, L. M., Aguilar, R. C., Naslavsky, N., Bonifacino, J. S. (2001). Human Vam6p promotes lysosome clustering and fusion in vivo. J. Cell Biol. 154: 109-122 [Abstract] [Full Text]  
• Huizing, M., Sarangarajan, R., Strovel, E., Zhao, Y., Gahl, W. A., Boissy, R. E. (2001). AP-3 Mediates Tyrosinase but Not TRP-1 Trafficking in Human Melanocytes. Mol. Biol. Cell 12: 2075-2085 [Abstract] [Full Text]  
• Kujala, P., Ikäheimonen, A., Ehsani, N., Vihinen, H., Auvinen, P., Kääriäinen, L. (2001). Biogenesis of the Semliki Forest Virus RNA Replication Complex. J. Virol. 75: 3873-3884 [Abstract] [Full Text]  
• Howard, T. L., Stauffer, D. R., Degnin, C. R., Hollenberg, S. M. (2001). CHMP1 functions as a member of a newly defined family of vesicle trafficking proteins. J. Cell Sci. 114: 2395-2404 [Abstract] [Full Text]  
• Rupper, A., Grove, B., Cardelli, J. (2001). Rab7 regulates phagosome maturation in Dictyostelium. J. Cell Sci. 114: 2449-2460 [Abstract] [Full Text]  
• Mullock, B. M., Smith, C. W., Ihrke, G., Bright, N. A., Lindsay, M., Parkinson, E. J., Brooks, D. A., Parton, R. G., James, D. E., Luzio, J. P., Piper, R. C. (2000). Syntaxin 7 Is Localized to Late Endosome Compartments, Associates with Vamp 8, and Is Required for Late Endosome-Lysosome Fusion. Mol. Biol. Cell 11: 3137-3153 [Abstract] [Full Text]  
• Pryor, P. R., Mullock, B. M., Bright, N. A., Gray, S. R., Luzio, J. P. (2000). The Role of Intraorganellar Ca2+ in Late Endosome-Lysosome Heterotypic Fusion and in the Reformation of Lysosomes from Hybrid Organelles. J. Cell Biol. 149: 1053-1062 [Abstract] [Full Text]  
• Blagoveshchenskaya, A. D., Cutler, D. F. (2000). Sorting to Synaptic-like Microvesicles from Early and Late Endosomes Requires Overlapping but Not Identical Targeting Signals. Mol. Biol. Cell 11: 1801-1814 [Abstract] [Full Text]  
• Bucci, C., Thomsen, P., Nicoziani, P., McCarthy, J., van Deurs, B. (2000). Rab7: A Key to Lysosome Biogenesis. Mol. Biol. Cell 11: 467-480 [Abstract] [Full Text]  
• Nicoziani, P., Vilhardt, F., Llorente, A., Hilout, L., Courtoy, P. J., Sandvig, K., van Deurs, B. (2000). Role for Dynamin in Late Endosome Dynamics and Trafficking of the Cation-independent Mannose 6-Phosphate Receptor. Mol. Biol. Cell 11: 481-495 [Abstract] [Full Text]  
• Reaves, B., Row, P., Bright, N., Luzio, J., Davidson, H. (2000). Loss of cation-independent mannose 6-phosphate receptor expression promotes the accumulation of lysobisphosphatidic acid in multilamellar bodies. J. Cell Sci. 113: 4099-4108 [Abstract]  
• Denzer, K, Kleijmeer, M., Heijnen, H., Stoorvogel, W, Geuze, H. (2000). Exosome: from internal vesicle of the multivesicular body to intercellular signaling device. J. Cell Sci. 113: 3365-3374 [Abstract]  
• Ohashi, M, Miwako, I, Yamamoto, A, Nagayama, K (2000). Arrested maturing multivesicular endosomes observed in a Chinese hamster ovary cell mutant, LEX2, isolated by repeated flow-cytometric cell sorting. J. Cell Sci. 113: 2187-2205 [Abstract]  
• Luzio, J., Rous, B., Bright, N., Pryor, P., Mullock, B., Piper, R. (2000). Lysosome-endosome fusion and lysosome biogenesis. J. Cell Sci. 113: 1515-1524 [Abstract]  
• Somsel Rodman, J, Wandinger-Ness, A (2000). Rab GTPases coordinate endocytosis. J. Cell Sci. 113: 183-192 [Abstract]  
• Bishop, N., Woodman, P. (2000). ATPase-defective Mammalian VPS4 Localizes to Aberrant Endosomes and Impairs Cholesterol Trafficking. Mol. Biol. Cell 11: 227-239 [Abstract] [Full Text]  
• Kundra, R., Kornfeld, S. (1999). Asparagine-linked Oligosaccharides Protect Lamp-1 and Lamp-2 from Intracellular Proteolysis. J. Biol. Chem. 274: 31039-31046 [Abstract] [Full Text]  
• Raposo, G., Cordonnier, M.-N., Tenza, D., Menichi, B., Dürrbach, A., Louvard, D., Coudrier, E. (1999). Association of Myosin I Alpha with Endosomes and Lysosomes in Mammalian Cells. Mol. Biol. Cell 10: 1477-1494 [Abstract] [Full Text]  
• Neufeld, E. B., Wastney, M., Patel, S., Suresh, S., Cooney, A. M., Dwyer, N. K., Roff, C. F., Ohno, K., Morris, J. A., Carstea, E. D., Incardona, J. P., Strauss, J. F. III, Vanier, M. T., Patterson, M. C., Brady, R. O., Pentchev, P. G., Blanchette-Mackie, E. J. (1999). The Niemann-Pick C1 Protein Resides in a Vesicular Compartment Linked to Retrograde Transport of Multiple Lysosomal Cargo. J. Biol. Chem. 274: 9627-9635 [Abstract] [Full Text]  
• Sunio, A., Metcalf, A. B., Krämer, H. (1999). Genetic Dissection of Endocytic Trafficking in Drosophila Using a Horseradish Peroxidase-Bride of Sevenless Chimera: hook Is Required for Normal Maturation of Multivesicular Endosomes. Mol. Biol. Cell 10: 847-859 [Abstract] [Full Text]  
• Coupin, G., Muller, C., Remy-Kristensen, A, Kuhry, J. (1999). Cell surface membrane homeostasis and intracellular membrane traffic balance in mouse L929 cells. J. Cell Sci. 112: 2431-2440 [Abstract]  
• Ohashi, M, Miwako, I, Nakamura, K, Yamamoto, A, Murata, M, Ohnishi, S., Nagayama, K (1999). An arrested late endosome-lysosome intermediate aggregate observed in a Chinese hamster ovary cell mutant isolated by novel three-step screening. J. Cell Sci. 112: 1125-1138 [Abstract]  
• Blagoveshchenskaya, A. D., Hewitt, E. W., Cutler, D. F. (1998). A Balance of Opposing Signals within the Cytoplasmic Tail Controls the Lysosomal Targeting of P-selectin. J. Biol. Chem. 273: 27896-27903 [Abstract] [Full Text]  
• Bryant, N. J., Piper, R. C., Weisman, L. S., Stevens, T. H. (1998). Retrograde Traffic Out of the Yeast Vacuole to the TGN Occurs via the Prevacuolar/Endosomal Compartment. J. Cell Biol. 142: 651-663 [Abstract] [Full Text]  
• Futter, C.E., Collinson, L.M., Backer, J.M., Hopkins, C.R. (2001). Human VPS34 is required for internal vesicle formation within multivesicular endosomes. J. Cell Biol. 155: 1251-1264 [Abstract] [Full Text]  

Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents