Endocytosis pp 113-121 | Cite as

Regulation of Endocytic Membrane Traffic

  • Jean Gruenberg
  • Jean-Pierre Gorvel
  • Morgane Bomsel
Conference paper
Part of the NATO ASI Series book series (volume 62)


Markers internalized into animal cells appear sequentially in peripheral early endosomes, then in late endosomes, predominantly located in the perinuclear region, and eventually in lysosomes (review, Gruenberg and Howell, 1989; Kornfeld and Mellman, 1989; see Fig 1). Our major interest is to understand the mechanisms regulating membrane traffic in this pathway. For these studies, we use a cell-free assay measuring endocytic vesicle fusion. Avidin and biotinylated horseradish peroxidase (bHRP) are separately internalized by fluid phase endocytosis into two cell populations. After homogenization, endosomal fractions are prepared by immuno-isolation (Gruenberg and Howell, 1986, 1987; Gruenberg et al., 1989) or flotation on gradients (Tuomikoski et al., 1989; Bomsel et al., 1989; Gorvel et al., submitted). In the assay, avidin- and bHRP-labeled fractions are combined with cytosol and incubated at 37°C in the presence of ATP and biotinylated insulin, as a scavenger (Braell, 1987; Gruenberg et al., 1989). When fusion occurs, a complex is formed between avidin and bHRP, which is then immuno-precipitated after detergent extraction. Fusion is quantified by measuring the enzymatic activity of bHRP in the complex. With this assay, we have reconstituted an early (Gruenberg et al., 1989) and a late (Bomsel et al., 1990) endocytic fusion event and we have shown that these reflect lateral interaction between early endosome elements and delivery to late endosomes, respectively.


Early Endosome Late Endosome Fusion Activity Lateral Fusion Cytoplasmic Dynein 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


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  1. Bacaliao, R, Antony, C., Dotti, C., Karsenti, E., Stelzer, E. H., and Simons, K. (1989). J. Cell Biol. 109, 2817–2832.CrossRefGoogle Scholar
  2. Beckers, C. J. M., Block, M. R, Glick, B. S., Rothman, J. E., and Balch, W. E. (1989). Nature 339, 397–398.PubMedCrossRefGoogle Scholar
  3. Block, M R., Glick, B. S., Wilcox, C. A., Wieland, F. T„ and Rothman, J.(1988) Proc. Natl. Acad. Sci. USA 85, 7852–7856.PubMedCrossRefGoogle Scholar
  4. Bomsel, M., Piydz, K., Parton, R. G., Gruenberg, J., and Simons, K. (1989). J. Cell Biol. 109, 3243–3258.PubMedCrossRefGoogle Scholar
  5. Bomsel, M., Parton, R., Kuznetsov, S. A., Schroer, T. A., and Gruenberg, J. (1990). Cell 62, 719–731.PubMedCrossRefGoogle Scholar
  6. Bourne, H. R. (1988). Cell 53, 669–671.PubMedCrossRefGoogle Scholar
  7. Braell, W. A. (1987). Proc. Natl. Acad. Sci. USA. 84, 1137–1141.Google Scholar
  8. Chavrier, P., Parton, R. G., Hauri, H. P., Simons, K., and Zerial, M. (1990). Cell 62, 317–329.PubMedCrossRefGoogle Scholar
  9. Davey, J., Hurtley, S. M., and Warren, G. (1985). Cell 43, 643–652.PubMedCrossRefGoogle Scholar
  10. De Brabander, M., Nuydens, R., Geerts, H., and Hopkins, C. R. (1988). Cell Mot. Cyt., 9, 30–47.CrossRefGoogle Scholar
  11. Deschenes, R. J., and Broach, J. R. (1987). Mol. Cell. Biol. 7, 2344–2351.Google Scholar
  12. Diaz, R., Mayorga, L., and Stahl, P. D. (1988). J. Biol. Chem. 263, 6093–6100.PubMedGoogle Scholar
  13. Diaz, R., Mayorga, L., Weidman, P. J., Rothman, J. E., and Stahl, P. D. (1988) Nature 339, 398–400.CrossRefGoogle Scholar
  14. Goda, Y., and Pfeffer, S. R. (1988). Cell 55, 309–320PubMedCrossRefGoogle Scholar
  15. Gorvel, J.-P., Chavrier, P., Zerial, M., and Gruenberg, J. (submitted).Google Scholar
  16. Griffiths, G., Back, R., and Marsh, M. (1989). J. Cell Biol. 109, 2703–2720.PubMedCrossRefGoogle Scholar
  17. Gruenberg, J., and Howell, K. E. (1986). EMBO J. 5, 3091–3101.PubMedGoogle Scholar
  18. Gruenberg, J., and Howell, K. E. (1987). Proc. Natl. Acad. Sci. USA 84, 5758–5762.PubMedCrossRefGoogle Scholar
  19. Gruenberg, J., and Howell, K. E. (1989). Annu. Rev. Cell Biol. 5, 453–481.PubMedCrossRefGoogle Scholar
  20. Gruenberg, J., Griffiths, G. and Howell, K. E. (1989). J. Cell Biol. 108, 1301–1316.PubMedCrossRefGoogle Scholar
  21. Herman, B. and Albertini, D. F. (1984). J. Cell Biol. 98, 565–576.PubMedCrossRefGoogle Scholar
  22. Hokins, C., Gibson, A., Shipman, M., and Miller, K. (1990). Nature, 346, 335–339.CrossRefGoogle Scholar
  23. Ingold, A. L., Cohn, S. A., and Scholey, J. M. (1988). J. Cell Biol. 107, 2657–2667.PubMedCrossRefGoogle Scholar
  24. Killisch, I., Steinlen, Römisch, K, Beug, H., and Griffiths, G. (submitted).Google Scholar
  25. Kornfeld, S., and Mellman, I. (1989). Annu. Rev. Cell Biol. 5, 483–525.PubMedCrossRefGoogle Scholar
  26. Labbé, J.-C., Capony, J.-P., Caput, D., Cavadore, J.-C., Derancourt, J., Kaghad, M., Lelias, J.-M., Picard, A. & Dorée, M. (1989). EMBO J. 8, 3053–3058.PubMedGoogle Scholar
  27. Malhotra, V., Orci, L., Glick, B. S., Block, M. R, and Rothman, J. E. (1988) Cell 54, 221–227.PubMedCrossRefGoogle Scholar
  28. Matteoni, R. and Kreis, T. E. (1987). J. Cell Biol. 105, 1253–1265.PubMedCrossRefGoogle Scholar
  29. Mayorga, L.S., Diaz, R., and Stahl, P. D. (1989). Science 244, 1475–1477.PubMedCrossRefGoogle Scholar
  30. Melançon, P., Glick, B. S., Malhotra, V., Weidman, P. J., Serafïni, T., Gleason, M. L., Orci, L., and Rothman, J. E. (1987). Cell 51, 1053–1062.PubMedCrossRefGoogle Scholar
  31. Molenaar, C. M. T., Prange, R., and Gallwitz, D. (1988). EMBO J. 7, 971–976.PubMedGoogle Scholar
  32. Mullock, B. M., Branch, W. J., van Schaik, M., Gilbert, M., and Luzio, J. P. (1988) J. Cell Biol. 108, 2093–2100.CrossRefGoogle Scholar
  33. Pai, E. F., Kabsh, W Krengel, U, Holmes, K. C, John, J, and Wittinghofer, A. (1989). Nature 341, 209–214.PubMedCrossRefGoogle Scholar
  34. Parton, R. G., Prydz, K, Bomsel, M., Simons, K., and Griffiths, G. (1989).J. Cell Biol. 109, 3259–3272.PubMedCrossRefGoogle Scholar
  35. Parton, R. G., Bacallao, R., and Piydz, K. (submitted). Schroer, T. A., Steuer, E. R., Sheetz, M. P. (1989). Cell, 56, 937–946.CrossRefGoogle Scholar
  36. Tuomikoski, T., Felix, M.-A., Dorée, M., and Gruenberg, J. (1989). Nature, 342, 942–945.PubMedCrossRefGoogle Scholar
  37. Warren, G. (1985). Trends Biochem. Sci. 503, 439–443.CrossRefGoogle Scholar
  38. Warren, G. (1989). Nature, 342, 857–858.PubMedCrossRefGoogle Scholar
  39. Walworth, N. C., Goud, B., Kastan Kabcenell, A., and Novick, P. J. (1989). EMBO J. 8, 1685–1693.PubMedGoogle Scholar
  40. Willumsen, B. M., Noms, K., Papageorge, A. G., Hubbert, N. L., and Lowy, D. R. (1984). EMBO J. 3, 2581–2585.PubMedGoogle Scholar
  41. Wilson, J. M., Whitney, J. A., and Neutra, M. R. (1987). J. Cell Biol. 105, 691–703.PubMedCrossRefGoogle Scholar
  42. Wilson, D. W., Wilcox, C. A., Flynn, G. C., Chen, E., Kuang, W.-J., Henzel, W. J., Block, M. R., Ullrich, A., and Rothman, J. E. (1989). Nature 339, 355–359.PubMedCrossRefGoogle Scholar
  43. Woodman, P. G., and Warren, G. (1988). Europ. J. Biochem. 173, 101–108PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1992

Authors and Affiliations

  • Jean Gruenberg
    • 2
  • Jean-Pierre Gorvel
    • 2
  • Morgane Bomsel
    • 1
  1. 1.Dpt of AnatomyUCSFSan FranciscoUSA
  2. 2.Europ. Mol. Biol. Lab.HeidelbergGermany

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