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Impairment of dynamin's GAP domain stimulates receptor-mediated endocytosis

Nature volume 398, pages 481–486 (1999)Cite this article

Abstract

Dynamin is a GTP-hydrolysing protein that is an essential participant in clathrin-mediated endocytosis by cells. It self-assembles into ‘collars’ in vitro which also form in vivo at the necks of invaginated coated pits. This self-assembly stimulates dynamin's GTPase activity and it has been proposed that dynamin hydrolyses GTP in order to generate the force needed to sever vesicles from the plasma membrane. A mechanism is now described in which self-assembly of dynamin is coordinated by a domain of dynamin with a GTPase-activating function. Unexpectedly, when dynamin mutants defective in self-assembly-stimulated GTPase activity are overexpressed, receptor-mediated endocytosis is accelerated. The results indicate that dynamin, like other members of the GTPase superfamily, functions as a molecular regulator in receptor-mediated endocytosis, rather than as a force-generating GTPase.

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Figure 1: GED stimulates both the minimal GTPase domain and full-length dynamin.
Figure 2: GED inhibits dynamin self-assembly and receptor-mediated endocytosis.
Figure 3: Identification of active residues within GED.
Figure 4: GTPase and self-assembly activities of full-length wild-type, R725A and K694A mutant dynamin
Figure 5: Receptor-mediated endocytosis is regulated by dynamin·GTP.
Figure 6: An intramolecular, assembly-dependent GAP functions to inactivate dynamin.

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Acknowledgements

We thank H. Damke for her contribution to analysis of dynamin function in vivo, L.M. Fujimoto and F. Simpson for advice and assistance, and J. Walter, W. E. Balch, L. Gerace, H. Damke and S. Newmyer for critically reading the manuscript. S.L.S. was supported by grants from the NIH and is an established investigator of the American Heart Association. S.S. was supported by a postdoctoral fellowship from the American Cancer Society and A.B.M. was supported by a predoctoral fellowship from the American Heart Association.

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  1. Department of Cell Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, California, 92037, La Jolla, USA

    Sanja Sever, Amy B. Muhlberg & Sandra L. Schmid

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  1. Sanja Sever
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Correspondence to Sandra L. Schmid.

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Sever, S., Muhlberg, A. & Schmid, S. Impairment of dynamin's GAP domain stimulates receptor-mediated endocytosis. Nature 398, 481–486 (1999). https://doi.org/10.1038/19024

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