Uncoupling of dynamin polymerization and GTPase activity revealed by the conformation-specific nanobody Dynab
- University of Geneva, Switzerland
- University of Valencia, Spain
- Institut Curie, France
Abstract
Dynamin is a large GTPase that forms a helical collar at the neck of endocytic pits, and catalyzes membrane fission (1, 2). Dynamin fission reaction is strictly dependent on GTP hydrolysis, but how fission is mediated is still debated (3): GTP energy could be spent in membrane constriction required for fission, or in disassembly of the dynamin polymer to trigger fission. To follow dynamin GTP hydrolysis at endocytic pits, we generated a conformation-specific nanobody called dynab, that binds preferentially to the GTP hydrolytic state of dynamin-1. Dynab allowed us to follow the GTPase activity of dynamin-1 in real-time. We show that in fibroblasts, dynamin GTP hydrolysis occurs as stochastic bursts, which are randomly distributed relatively to the peak of dynamin assembly. Thus, dynamin disassembly is not coupled to GTPase activity, supporting that the GTP energy is primarily spent in constriction.
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Author details
Funding
Human Frontier Science Program (CDA-0061-08)
- Aurélien Roux
Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (Grant 31003A_130520)
- Aurélien Roux
H2020 European Research Council (Starting Grant 311536 (2011 call))
- Aurélien Roux
Seventh Framework Programme (Marie Curie ITN grant #264399)
- Aurélien Roux
Agence Nationale de la Recherche (ANR-12-BSV2-0003-01)
- Franck Perez
Centre National de la Recherche Scientifique
- Franck Perez
Fondation pour la Recherche Médicale (DEQ20120323723)
- Franck Perez
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2017, Galli et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Uncoupling of dynamin polymerization and GTPase activity revealed by the conformation-specific nanobody Dynab
eLife 6:e25197.
https://doi.org/10.7554/eLife.25197
