WAVE regulatory complex activation by cooperating GTPases Arf and Rac1

Koronakis, Vassilis; Hume, Peter; Humphreys, Daniel; Liu, Tao; Hørning, Ole; Jensen, Ole N.; McGhie, Emma J.

doi:10.1073/pnas.1107666108

Cited by 126 publications

(187 citation statements)

References 35 publications

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“…Here, we establish that Arf79F not only localises at the cell edge with the WRC component Sra1 but is also essential for lamellipodium formation. This builds on our recent demonstration that Arf1 directly recruits and activates mammalian WRC in vitro (Koronakis et al, 2011).…”

Section: Resultsmentioning

confidence: 68%

“…We recently demonstrated that mammalian Arf1 directly binds the structural homologues Sra1 and Nap1 (Koronakis et al, 2011) and therefore wondered whether Arf79F drives lamellipodium formation by recruiting WRC to the plasma membrane. First, purified in vitro myristoylated Arf79F was anchored to silica beads coated in a phospholipid bilayer consisting of equal amounts of phosphatidylcholine and phosphatidylinositol (PC:PI).…”

Section: Resultsmentioning

confidence: 99%

“…This role appears to be specific for Arf79F as Class 1 but not Class 2 or 3 Arfs were required for lamellipodia. This was surprising as WRC can be activated in cell extract by divergent Arf GTPases such as Arf5 (Arf102F homologue) and Arl1 (Koronakis et al, 2011). However, Gautier and colleagues showed that Arf79F, but not related Drosophila Arf isoforms, was found bound to immunoprecipitated WRC underlining Arf specificity for the WRC.…”

Section: Resultsmentioning

confidence: 99%

“…However, Gautier and colleagues showed that Arf79F, but not related Drosophila Arf isoforms, was found bound to immunoprecipitated WRC underlining Arf specificity for the WRC. Since Arf1 alone is insufficient for WRC activation in vitro (Koronakis et al, 2011) and Arf must cooperate with Rac1 for WRC-driven actin assembly (Humphreys et al, 2012;Koronakis et al, 2011), it seems that it is only active Arf79F which is temporally and spatially coincident with Rac1 activation in Drosophila cells.…”

Section: Resultsmentioning

confidence: 99%

“…The low affinity of Rac1 for WRC (Chen et al, 2010) suggested that additional unknown factors are required to facilitate WRC-dependent actin polymerisation (Davidson and Insall, 2011). We showed that active Rac1 was indeed insufficient for recruitment and activation of the WRC at the membrane in mammalian cell extract (Koronakis et al, 2011). This led to our discovery that Arf family GTPases directly recruit and activate the WRC, which is enhanced when Arf and Rac1 work together to trigger actin polymerisation.…”

Section: Introductionmentioning

confidence: 97%

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The Drosophila Arf1 homologue Arf79F is essential for lamellipodium formation

Humphreys

Liu

Davidson

et al. 2012

Journal of Cell Science

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SummaryThe WAVE regulatory complex (WRC) drives the polymerisation of actin filaments located beneath the plasma membrane to generate lamellipodia that are pivotal to cell architecture and movement. By reconstituting WRC-dependent actin assembly at the membrane, we recently discovered that several classes of Arf family GTPases directly recruit and activate WRC in cell extracts, and that Arf cooperates with Rac1 to trigger actin polymerisation. Here, we demonstrate that the Class 1 Arf1 homologue Arf79F colocalises with the WRC at dynamic lamellipodia. We report that Arf79F is required for lamellipodium formation in Drosophila S2R+ cells, which only express one Arf isoform for each class. Impeding Arf function either by dominant-negative Arf expression or by Arf doublestranded RNA interference (dsRNAi)-mediated knockdown uncovered that Arf-dependent lamellipodium formation was specific to Arf79F, establishing that Class 1 Arfs, but not Class 2 or Class 3 Arfs, are crucial for lamellipodia. Lamellipodium formation in Arf79F-silenced cells was restored by expressing mammalian Arf1, but not by constitutively active Rac1, showing that Arf79F does not act via Rac1. Abolition of lamellipodium formation in Arf79F-silenced cells was not due to Golgi disruption. Blocking Arf79F activation with guanine nucleotide exchange factor inhibitors impaired WRC localisation to the plasma membrane and concomitant generation of lamellipodia. Our data indicate that the Class I Arf GTPase is a central component in WRC-driven lamellipodium formation.

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Section: Resultsmentioning

confidence: 68%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 97%

See 3 more Smart Citations

The Drosophila Arf1 homologue Arf79F is essential for lamellipodium formation

Humphreys

Liu

Davidson

et al. 2012

Journal of Cell Science

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The Arf family GTPases: Regulation of vesicle biogenesis and beyond

Guan

2023

BioEssays

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The Arf family proteins are best known for their roles in the vesicle biogenesis. However, they also play fundamental roles in a wide range of cellular regulation besides vesicular trafficking, such as modulation of lipid metabolic enzymes, cytoskeleton remodeling, ciliogenesis, lysosomal, and mitochondrial morphology and functions. Growing studies continue to expand the downstream effector landscape of Arf proteins, especially for the less‐studied members, revealing new biological functions, such as amino acid sensing. Experiments with cutting‐edge technologies and in vivo functional studies in the last decade help to provide a more comprehensive view of Arf family functions. In this review, we summarize the cellular functions that are regulated by at least two different Arf members with an emphasis on those beyond vesicle biogenesis.

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Structural features and interfacial properties of WH2, β‐thymosin domains and other intrinsically disordered domains in the regulation of actin cytoskeleton dynamics

2013

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Many actin-binding proteins (ABPs) use complex multidomain architectures to integrate and coordinate multiple signals and interactions with the dynamic remodeling of actin cytoskeleton. In these proteins, small segments that are intrinsically disordered in their unbound native state can be functionally as important as identifiable folded units. These functional intrinsically disordered regions (IDRs) are however difficult to identify and characterize in vitro. Here, we try to summarize the state of the art in understanding the structural features and interfacial properties of IDRs involved in actin self-assembly dynamics. Recent structural and functional insights into the regulation of widespread, multifunctional WH2/b-thymosin domains, and of other IDRs such as those associated with WASP/ WAVE, formin or capping proteins are examined. Understanding the functional versatility of IDRs in actin assembly requires apprehending by multiple structural and functional approaches their large conformational plasticity and dynamics in their interactions. In many modular ABPs, IDRs relay labile interactions with multiple partners and act as interaction hubs in interdomain and protein-protein interfaces. They thus control multiple conformational transitions between the inactive and active states or between various active states of multidomain ABPs, and play an important role to coordinate the high turnover of interactions in actin self-assembly dynamics. V C 2013 Wiley Periodicals, Inc.

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WAVE regulatory complex activation by cooperating GTPases Arf and Rac1

Cited by 126 publications

References 35 publications

The Drosophila Arf1 homologue Arf79F is essential for lamellipodium formation

The Drosophila Arf1 homologue Arf79F is essential for lamellipodium formation

The Arf family GTPases: Regulation of vesicle biogenesis and beyond

Structural features and interfacial properties of WH2, β‐thymosin domains and other intrinsically disordered domains in the regulation of actin cytoskeleton dynamics

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