WAVE binds Ena/VASP for enhanced Arp2/3 complex–based actin assembly

Havrylenko, Svitlana; Noguera, Philippe; Abou-Ghali, Majdouline; Manzi, John; Faqir, Fahima; Lamora, Audrey; Guérin, Christophe; Blanchoin, Laurent; Plastino, Julie

doi:10.1091/mbc.e14-07-1200

Cited by 63 publications

(65 citation statements)

References 74 publications

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“…The authors concluded that profilin antagonizes Arp2/3-mediated nucleation and actin branching by facilitating actin polymerization via Ena/VASP, thereby competing away actin monomers from the Arp2/3 complex. In contrast, Ena/VASP has also been proposed to interact with the WAVE regulatory complex to stimulate Arp2/3 activation, enhance migration, and maintain normal lamellipodia formation in Drosophila hemocytes and C. elegans epidermal cells, suggesting that this family of actin assembly proteins may have alternative functions in diverse cell types or under different conditions [106,107]. Work combining analysis of the contractile ring in S. pombe with in vitro actin reconstitution assays also shows that profilin inhibits Arp2/3 function, thereby maintaining the balance between formin- and Arp2/3-mediated actin polymerization that is required for cytokinesis [108].…”

Section: Introductionmentioning

confidence: 99%

Function and regulation of the Arp2/3 complex during cell migration in diverse environments

Swaney

2016

Current Opinion in Cell Biology

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As the first de novo actin nucleator discovered, the Arp2/3 complex has been a central player in models of protrusive force production via the dynamic actin network. Here, we review recent studies on the functional role of the Arp2/3 complex in the migration of diverse cell types in different migratory environments. These findings have revealed an unexpected level of plasticity, both in how cells rely on the Arp2/3 complex for migration and other physiological functions and in the intricate modulation of the Arp2/3 complex by other actin regulators and upstream signaling cascades.

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Section: Introductionmentioning

confidence: 99%

Function and regulation of the Arp2/3 complex during cell migration in diverse environments

Swaney

2016

Current Opinion in Cell Biology

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“…So the effect of VASP on motility seems to depend on the initial state of the system, and when speed is already optimal, adding an enhancing molecule like VASP does not have the expected effect. Images taken at about 10-15 minutes reaction time of PRD-VCA-WAVE-coated beads in reconstituted motility mix as described in [65], but with Arp2/3 complex purchased from Cytoskeleton. Phase contrast microscopy.…”

Section: Resultsmentioning

confidence: 99%

“…N-WASP is a more effective Arp2/3 complex activator than either WASP or WAVE/Scar due to the enhanced acidity of the A domain in the case of N-WASP, not as originally believed due to the extra V domain that N-WASP proteins contain [64]. WAVE/Scar-derived bead coatings have been used for some studies, but less extensively than the other NPFs [46,65]. WAVE proteins exist in regulatory complexes, which are impossible to mimic in pure protein mixtures although the WAVE regulatory complex has been successfully recruited to membrane-coated glass beads to form actin comets in cell extracts [66].…”

Section: What To Coat the Beads With?mentioning

confidence: 99%

“…As one example, this approach was used to resolve the confusion concerning Ena/VASP proteins and Listeria motility mentioned previously. When recruited to the bead surface, Ena/VASP proteins were shown to indeed increase bead speed and different mutants of Ena/VASP showed concordant effects on beads and on an in vivo cell motility event [49,65,72]. However the relation between actin polymerization and particle speed is a complex one.…”

Section: The Power Of the Bead System In The Pure Protein MIXmentioning

confidence: 99%

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Reconstituting the actin cytoskeleton at or near surfaces in vitro

Cáceres

Abou-Ghali

Plastino

2015

Biochimica et Biophysica Acta (BBA) - Molecular Cell Research

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Actin filament dynamics have been studied for decades in pure protein solutions or in cell extracts, but a breakthrough in the field occurred at the turn of the century when it became possible to reconstitute networks of actin filaments, growing in a controlled but physiological manner on surfaces, mimicking the actin assembly that occurs at the plasma membrane during cell protrusion and cell shape changes. The story begins with the bacteria Listeria monocytogenes, the study of which led to the reconstitution of cellular actin polymerization on a variety of supports including plastic beads. These studies made possible the development of liposome-type substrates for filament assembly and micropatterning of actin polymerization nucleation. Based on the accumulated expertise of the last 15 years, many exciting approaches are being developed, including the addition of myosin to biomimetic actin networks to study the interplay between actin structure and contractility. The field is now poised to make artificial cells with a physiological and dynamic actin cytoskeleton, and subsequently to put these cells together to make in vitro tissues. This article is part of a Special Issue entitled: Mechanobiology.

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“…The EVH1 domains target Ena/VASP proteins to sites of actin rearrangements via interactions with proteins containing the proline-rich motif (D/E)(F/L/W/Y)PPPPX(D/E)(D/E) (designated FPPPP), including zyxin and vinculin at focal adhesions and lamellipodin at the leading edge (Hoffman et al, 2006;Krause et al, 2004;Niebuhr et al, 1997). EVH1 domains also bind to diaphanous formin (Dia) and mammalian diaphanous formin 2 (mDia2), inhibiting actin nucleation and elongation (Barzik et al, 2014;Bilancia et al, 2014), and recent data suggest that VASP activates WAVE (Wiskott-Aldrich syndrome verprolin homology protein) via EVH1-mediated interactions (Chen et al, 2014;Havrylenko et al, 2015). The PRO regions bind to profilin and to SH3 and WW domain-containing proteins.…”

Section: Introductionmentioning

confidence: 99%

Vasodilator-stimulated phosphoprotein restricts cell-to-cell spread of Shigella flexneri at the cell periphery

2015

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Shigella spp. are intracellular bacterial pathogens that cause diarrhoeal disease in humans. Shigella utilize the host actin cytoskeleton to enter cells, move through the cytoplasm of cells and pass into adjacent cells. Ena/VASP family proteins are highly conserved proteins that participate in actin-dependent dynamic cellular processes. We tested whether Ena/VASP family members VASP (vasodilator-stimulated phosphoprotein), Mena (mammalian-enabled) or EVL (Ena-VASP-like) contribute to Shigella flexneri spread through cell monolayers. VASP and EVL restricted cell-to-cell spread without significantly altering actin-based motility, whereas Mena had no effect on these processes. Phosphorylation of VASP on Ser153, Ser235 and Thr274 regulated its subcellular distribution and function. VASP derivatives that lack the Ena/VASP homology 1 (EVH1) domain or contain a phosphoablative mutation of Ser153 were defective in restricting S. flexneri spread, indicating that the EVH1 domain and phosphorylation on Ser153 are required for this process. The EVH1 domain and Ser153 of VASP were required for VASP localization to focal adhesions, and localization of VASP to focal adhesions and/or the leading edge was required for restriction of spread. The contribution of the EVH1 domain was from both the donor and the recipient cell, whereas the contribution of Ser153 phosphorylation was only from the donor cell. Thus, unlike host proteins characterized in Shigella pathogenesis that promote bacterial spread, VASP and EVL function to limit it. The ability of VASP and EVL to limit spread highlights the critical role of focal adhesion complexes and/or the leading edge in bacterial passage between cells.

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WAVE binds Ena/VASP for enhanced Arp2/3 complex–based actin assembly

Abstract: A dual in vitro/in vivo approach is used to show that WAVE directly binds Ena/VASP, coordinating its activity with that of the Arp2/3 complex for enhanced actin assembly.

Cited by 63 publications

References 74 publications

Function and regulation of the Arp2/3 complex during cell migration in diverse environments

Function and regulation of the Arp2/3 complex during cell migration in diverse environments

Reconstituting the actin cytoskeleton at or near surfaces in vitro

Vasodilator-stimulated phosphoprotein restricts cell-to-cell spread of Shigella flexneri at the cell periphery

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