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The crystal structure of the inactive form of the Arp2/3 complex is described. The structure defines ARPC2 and ARPC4 as the structural core of the complex. The location and polarity of Arp2 and Arp3 are consistent with their serving as the nucleation site for a filament with a free barbed end, but a substantial change in the position of the subunits relative to each other probably accompanies activation.
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The structure of the Arp2/3 complex at actin filament branches is described from cryo-electron microscopy. The Arp2/3 complex binds along the side of the mother filament. Activation of the Arp2/3 complex appears to involve large structural changes.
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CARMIL acts as a scaffold, binding myosin-I, Arp2/3 complex, and capping protein. A GST fusion protein containing an acidic stretch of CARMIL (residues 644-863) weakly activates Arp2/3-dependent actin nucleation. The localization and mutant phenotype demonstrate an important role for CARMIL in actin-based cell motility.
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WAVE proteins are negatively regulated by associated proteins. Rac and Nck relieve this inhibition, apparently by dissociating the inhibitor proteins.
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•] demonstrate that cortactin, a Src kinase substrate and F-actin-binding protein, activates the actin nucleation activity of Arp2/3 complex. Weaver et al. also show that cortactin stabilises the actin filament branch points formed by the Arp2/3 complex.
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•] demonstrate that cortactin, a Src kinase substrate and F-actin-binding protein, activates the actin nucleation activity of Arp2/3 complex. Weaver et al. also show that cortactin stabilises the actin filament branch points formed by the Arp2/3 complex.
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WIP interacts with N-WASP to control Arp2/3-mediated actin assembly in vitro and in vivo. WIP and N-WASP together are important for the formation of filopodia by Cdc42 and bradykinin.
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Cortactin and neuronal Wiskott-Aldrich syndrome protein (N-WASP) can bind simultaneously to Arp2/3 complex, providing synergistic stimulation of actin assembly. They share a binding site on Arp3, and N-WASP has additional interactions with Arp2 and ARPC1/p40. Interaction of the DDW motif of N-WASP with Arp3 is not important for activation of Arp2/3 complex.
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Madania A., Dumoulin P., Grava S., Kitamoto H., Scharer-Brodbeck C., Soulard A., Moreau V., Winsor B. The Saccharomyces cerevisiae homologue of human Wiskott-Aldrich syndrome protein Las17p interacts with the Arp2/3 complex. Mol. Biol. Cell. 10:1999;3521-3538.
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Yeast Abp1p binds and activates the actin nucleation activity of Arp2/3 complex. The mechanism is similar to that of cortactin, increasing the association of Arp2/3 complex with filamentous actin.
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Association of mouse actin-binding protein 1 (mAbp1/sh3p7), an Src kinase target, with dynamic regions of the cortical actin cytoskeleton in response to Rac1 activation
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Myo1p and Wsp1p, the fission yeast WASP-like protein, share functions and cooperate in controlling actin assembly. A GST fusion protein containing the actin-filament-binding TH2 domain and the A domain of the Myo1p tail binds to the Arp2/3 complex and activates its actin nucleation activity.
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Pan1p of budding yeast activates the actin nucleation activity of the Arp2/3 complex and also interacts with several clathrin-associated proteins involved in endocytosis. Pan1p can provide a biochemical link between the endocytic machinery and actin assembly.
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