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Arp2/3 ATP hydrolysis-catalysed branch dissociation is critical for endocytic force generation

Nature Cell Biology volume 8, pages 826–833 (2006)Cite this article

Abstract

The Arp2/3 complex, which is crucial for actin-based motility, nucleates actin filaments and organizes them into y-branched networks. The Arp2 subunit has been shown to hydrolyse ATP, but the functional importance of Arp2/3 ATP hydrolysis is not known. Here, we analysed an Arp2 mutant in Saccharomyces cerevisiae that is defective in ATP hydrolysis. Arp2 ATP hydrolysis and Arp2/3-dependent actin nucleation occur almost simultaneously. However, ATP hydrolysis is not required for nucleation. In addition, Arp2 ATP hydrolysis is not required for the release of a WASP-like activator from y-branches. ATP hydrolysis by Arp2, and possibly Arp3, is essential for efficient y-branch dissociation in vitro. In living cells, both Arp2 and Arp3 ATP-hydrolysis mutants exhibit defects in endocytic internalization and actin-network disassembly. Our results suggest a critical feature of dendritic nucleation in which debranching and subsequent actin-filament remodelling and/or depolymerization are important for endocytic vesicle morphogenesis.

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Figure 1: Arp2 ATP hydrolysis is not required for actin filament nucleation.
Figure 2: Arp2 ATP hydrolysis is required for productive endocytic internalization.
Figure 3: Arp2 ATP hydrolysis is required for proper actin network disassembly.
Figure 4: Arp2, and possibly Arp3, ATP hydrolysis is required for efficient y-branch dissociation.
Figure 5: Schematic representation of a model for Arp2/3 complex ATP hydrolysis function.

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Acknowledgements

We are very grateful to C. Toret and M. Kaksonen for providing yeast strains used in this study, P. Carlton for help with the calculation of branching frequency, and to S. Almo for helpful suggestions about potentially informative Arp2/3 mutations. In addition, we thank E. Goley, M. Kaksonen, V. Okreglak, C. Toret, Y. Sun, J. Wong, B. Pauly and A. Engqvist-Goldstein for their comments on the manuscript. Finally, we thank members of both the Drubin and Welch labs for their technical help and encouragement. This work was supported by National Institute of Health (NIH) grants GM42759 and GM50399 to D.G.D. and NIH grant GM59609 to M.D.W.

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  1. 16 Barker Hall, Dept. of Molecular and Cell Biology, University of California, Berkeley, 94720-3202, CA, USA

    Adam C. Martin, Matthew D. Welch & David G. Drubin

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Correspondence to David G. Drubin.

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Martin, A., Welch, M. & Drubin, D. Arp2/3 ATP hydrolysis-catalysed branch dissociation is critical for endocytic force generation. Nat Cell Biol 8, 826–833 (2006). https://doi.org/10.1038/ncb1443

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