Abstract
Dynamin is a large GTPase with a relative molecular mass of 96,000 (Mr 96K) that is involved in clathrin-mediated endocytosis and other vesicular trafficking processes1,2. Although its function is apparently essential for scission of newly formed vesicles from the plasma membrane, the nature of dynamin's role in the scission process is still unclear3,4. It has been proposed that dynamin is a regulator (similar to classical G proteins) of downstream effectors5. Here we report the analysis of several point mutants of dynamin's GTPase effector (GED) and GTPase domains. We show that oligomerization and GTP binding alone, by dynamin, are not sufficient for endocytosis in vivo. Rather, efficient GTP hydrolysis and an associated conformational change are also required. These data argue that dynamin has a mechanochemical function in vesicle scission.
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References
Urrutia, R., Henley, J. R., Cook, T. & McNiven, M. A. The dynamins: redundant or distinct functions for an expanding family of related GTPases? Proc. Natl Acad. Sci. USA 94, 377–384 (1997).
van der Bliek, A. M. Functional diversity in the dynamin family. Trends Cell Biol. 9, 96–102 (1999).
Warnock, D. E., HInshaw, J. E. & Schmid, S. L. Dynamin self-assembly stimulates its GTPase activity. J. Biol. Chem. 271, 22310–22314 (1996).
Roos, J. & Kelly, R. B. Is dynamin really a ‘pinchase’? Trends Cell Biol. 7, 257–259 (1997).
Sever, S., Muhlberg, A. B. & Schmid, S. L. Impairment of dynamin's GAP domain stimulates receptor-mediated endocytosis. Nature 398, 481–486 (1999).
Poodry, C. A. & Edgar, L. Reversible alterations in the neuromuscular junctions of Drosophila melanogaster bearing a temperature-sensitive mutation, shibire. J. Cell Biol. 81, 520–527 (1979).
Kosaka, T. & Ikeda, K. Possible temperature-dependent blockage of synaptic vesicle recycling induced by a single gene mutation in Drosophila. J. Neurobiol. 14, 207–225 (1983).
Koenig, J. H. & Ikeda, K. Disappearance and reformation of synaptic vesicle membrane upon transmitter release observed under reversible blockage of membrane retrieval. J. Neurosci. 9, 3844–3860 (1989).
van der Bliek, A. M. et al. Mutations in human dynamin block an intermediate stage in coated vesicle formation. J. Cell Biol. 122, 553–563 (1993).
Herskovits, J. S., Burgess, C. C., Obar, R. A. & Vallee, R. B. Effects of mutant rat dynamin on endocytosis. J. Cell Biol. 122, 565–578 (1993).
Damke, H., Baba, T., Warnock, D. E. & Schmid, S. L. Induction of mutant dynamin specifically blocks endocytic coated vesicle formation. J. Cell Biol. 127, 915–934 (1994).
Takei, K., McPherson, P. S., Schmid, S. L. & DeCamilli, P. Tubular membrane invaginations coated by dynamin rings are induced by GTPγS in nerve terminals. Nature 374, 186–190 (1995).
Sweitzer, S. M. & Hinshaw, J. E. Dynamin undergoes a GTP-dependent conformational change causing vesiculation. Cell 93, 1021–1029 (1998).
Takei, K. et al. Generation of coated intermediates of clathrin-mediated endocytosis on protein-free liposomes. Cell 94, 131–141 (1998).
Stowell, M. H. B., Marks, B., Wigge, P. & McMahon, H. T. Nucleotide-dependent conformational changes in dynamin: evidence for a mechanochemical molecular spring. Nature Cell Biology 1, 27–32 (1999).
Takei, K., Slepnev, V. I., Haucke, V. & De Camilli, P. Functional partnership between amphiphysin and dynamin in clathrin-mediated endocytosis. Nature Cell Bio. 1, 33–39 (1999).
Muhlberg, A. B., Warnock, D. E. & Schmid, S. L. Domain structure and intramolecular regulation of dynamin GTPase. EMBO J. 16, 6676–6683 (1998).
Prakash, B., Renault, L., Praefcke, G. J., Herrmann, C. & Wittinghofer, A. Triphosphate structure of guanylate-binding protein 1 and implications for nucleotide binding and GTPase mechanism. EMBO J. 19, 4555–4564 (2000).
Okamoto, P. M., Tripet, B., Litowski, J., Hodges, R. S. & Vallee, R. B. Multiple distinct coiled-coils are involved in dynamin self-assembly. J. Biol. Chem. 274, 10277–10286 (1999).
Pai, E. F. et al. Refined crystal structure of the triphosphate conformation of H-ras p21 at 1.35 A resolution: implications for the mechanism of GTP hydrolysis. EMBO J. 9, 2351–2359 (1990).
Otero, A. D. Transphosphorylation and G protein activation. Biochem. Pharmacol. 39, 1399–1404 (1990).
Wilson-Kubalek, E. M., Brown, R. E., Celia, H. & Milligan, R. A. Lipid nanotubes as substrates for helical crystallization of macromolecules. Proc. Natl Acad. Sci. USA 95, 8040–8050 (1998).
Hopkins, C. R. & Trowbridge, I. S. Internalization and processing of transferrin and the transferrin receptor in human carcinoma A431 cells. J. Cell Biol. 97, 508–521 (1983).
Vallis, Y., Wigge, P., Marks, B., Evans, P. R. & McMahon, H. T. Importance of the pleckstrin homology domain of dynamin in clathrin-mediated endocytosis. Curr. Biol. 9, 257–260 (1999).
Achiriloaie, M., Barylko, B. & Albanesi, J. P. Essential role of the dynamin pleckstrin homology domain in receptor mediated endocytosis. Mol. Cell. Biol. 19, 1410–1415 (1999).
Smirnova, E., Shurland, D. L., Newman-Smith, E. D., Pishvaee, B. & van der Bliek, A. M. A model for dynamin self-assembly based on binding between three different protein domains. J. Biol. Chem. 274, 14942–14947 (1999).
Grabs, D. et al. The SH3 domain of amphiphysin binds the proline-rich domain of dynamin at a single site that defines a new SH3 binding consensus sequence. J. Biol. Chem. 272, 13419–13425 (1997).
Wigge, P. et al. Amphiphysin heterodimers: potential role in clathrin-mediated endocytosis. Mol. Biol. Cell 8, 2003–2015 (1997).
Owen, D. J. et al. Crystal structure of the Amphiphysin-2 SH3 domain and its role in prevention of dynamin ring formation. EMBO J. 17, 5273–5285 (1998).
Acknowledgements
We thank P. Wigge for discussions, M. Ford for discussions and assistance with immunofluorescence work, and M. Higgins for help with the expression of dynamin mutants.
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Marks, B., Stowell, M., Vallis, Y. et al. GTPase activity of dynamin and resulting conformation change are essential for endocytosis. Nature 410, 231–235 (2001). https://doi.org/10.1038/35065645
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DOI: https://doi.org/10.1038/35065645
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