Focal Adhesions Undergo Longitudinal Splitting into Fixed-Width Units

Young, Lorna E.; Higgs, Henry N.

doi:10.1016/j.cub.2018.04.073

Cited by 33 publications

(31 citation statements)

References 52 publications

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“…One possibility is that VASP and Mena might bind barbed-ends at different times during filopodial assembly, or that one of the proteins is indirectly linked to filopodia. For example, we recently showed that VASP plays a role in the FA 'splitting' process, whereby FAs longitudinally divide into 300 nm width units (Young and Higgs, 2018). Alternately, VASP and Mena might work together in the same process, with specific functions for each.…”

Section: Discussionmentioning

confidence: 99%

“…We next examined whether VASP and FMNL3 show similar filopodial localization patterns in actively spreading cells, using a collagen-based system we recently developed (Young and Higgs, 2018). Here, cells spread rapidly and uniformly in the first hour after plating, and collagen-plated cells typically display higher filopodial density during spreading ( Fig.…”

Section: Vasp Depletion Alters the Filopodia-adhesion Relationship Dumentioning

confidence: 99%

“…Ena/VASP proteins display clear filopodial tip enrichment in some situations (Applewhite et al, 2007;Bear et al, 2002;Disanza et al, 2013;Lanier et al, 1999;Reinhard et al, 1992;Rottner et al, 1999;Schäfer et al, 2009;Schirenbeck et al, 2006), but are absent from >90% of filopodia generated upon the overexpression of the constitutively active mDia2 (Barzik et al, 2014). VASP also enriches in focal adhesions (FAs), where it plays roles in dorsal stress fiber assembly (Gateva et al, 2014;Tojkander et al, 2015) and focal adhesion dynamics (Young and Higgs, 2018). For the formin family, mDia2, FMNL2 and FMNL3 have been shown to enrich at filopodial tips (Block et al, 2012(Block et al, , 2008Gauvin et al, 2015;Pellegrin and Mellor, 2005;Yang et al, 2007;Young et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

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Roles for Ena/VASP proteins in FMNL3-mediated filopodial assembly

Young

Latario

Higgs

2018

Journal of Cell Science

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Filopodia are actin-dependent finger-like structures that protrude from the plasma membrane. Actin filament barbed-end-binding proteins localized to filopodial tips are key to filopodial assembly. Two classes of barbed-end-binding proteins are formins and Ena/VASP proteins, and both classes have been localized to filopodial tips in specific cellular contexts. Here, we examine the filopodial roles of the FMNL formins and Ena/VASP proteins in U2OS cells. FMNL3 suppression reduces filopodial assembly by 90%, and FMNL3 is enriched at >95% of filopodial tips. Suppression of VASP or Mena (also known as ENAH) reduces filopodial assembly by >75%. However, VASP and Mena do not display consistent filopodial tip localization, but are enriched in focal adhesions (FAs). Interestingly, >85% of FMNL3containing filopodia are associated with FAs. Two situations increase Ena/VASP filopodial localization: (1) expression of myosin-X, and (2) actively spreading cells. In spreading cells, filopodia often mark sites of nascent adhesions. Interestingly, VASP suppression in spreading cells causes a significant increase in adhesion assembly at filopodial tips. This work demonstrates that, in U2OS cells, Ena/VASP proteins play roles in filopodia beyond those at filopodial tips. This article has an associated First Person interview with the first author of the paper.

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Section: Discussionmentioning

confidence: 99%

Section: Vasp Depletion Alters the Filopodia-adhesion Relationship Dumentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Roles for Ena/VASP proteins in FMNL3-mediated filopodial assembly

Young

Latario

Higgs

2018

Journal of Cell Science

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“…Finally, although Ena/VASP proteins are prominent FA components, their roles in cell substrate adhesion and FA formation have also remained controversial. Both inhibitory (Galler et al, 2006) and stimulatory roles (Gupton et al, 2012; Kang et al, 2010; Puleo et al, 2019; Young and Higgs, 2018) on adhesion were reported, whereas one study did not detect any effect in fibroblasts lacking Mena and VASP (Bear et al, 2000). To resolve these discrepancies, we characterized cells disrupted for individual, two or all three Ena/VASP family members in distinct types of parental cells, and explored the impact of their individual and collective removal on formation of different protrusion types, migration efficiency and adhesion.…”

Section: Introductionmentioning

confidence: 99%

Loss of Ena/VASP interferes with lamellipodium architecture, motility and integrin-dependent adhesion

Damiano-Guercio

Kurzawa

Mueller

et al. 2020

Preprint

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AbstractCell migration entails networks and bundles of actin filaments termed lamellipodia and microspikes or filopodia, respectively, as well as focal adhesions, all of which recruit Ena/VASP family members hitherto thought to antagonize efficient cell motility. However, we find these proteins to act as positive regulators of migration in different cell lines. CRISPR/Cas9-mediated loss of Ena/VASP proteins reduced lamellipodial actin assembly and perturbed lamellipodial architecture, as evidenced by changed network geometry as well as reduction of filament length and number that was accompanied by abnormal Arp2/3 complex and heterodimeric capping protein accumulation. Loss of Ena/VASP function also abolished the formation of microspikes normally embedded in lamellipodia, but not of filopodia capable of emanating without lamellipodia. Ena/VASP-deficiency also impaired integrin-mediated adhesion accompanied by reduced traction forces exerted through these structures. Our data thus uncover novel, cellular Ena/VASP functions of these actin polymerases that are fully consistent with their promotion of cell migration.

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“…While both integrin receptors themselves and numerous proteins linking the cytoplasmic domains of these receptors with the actin cytoskeleton are similar in podosomes and focal adhesions, the organization of the actin core is different in these types of structures. Focal adhesions are peripheral termini of actin bundles known as stress fibres and the actin scaffold of focal adhesions also consists of bundles of parallel actin filaments [13][14][15]. Actin filaments of focal adhesions are associated via numerous links containing talin and vinculin with clusters of integrins located immediately underneath the actin filament layer [16].…”

Section: Introductionmentioning

confidence: 99%

Forces and constraints controlling podosome assembly and disassembly

Rafiq

Grenci

Kozlov

et al. 2018

Preprint

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Podosomes are a singular category of integrin-mediated adhesions important in the processes of cell migration, matrix degradation, and cancer cell invasion.Despite a wealth of biochemical studies, the effects of mechanical forces on podosome integrity and dynamics are poorly understood. Here, we show that podosomes are highly sensitive to two groups of physical factors. First, we describe the process of podosome disassembly induced by activation of myosin-IIA filament assembly. Next, we find that podosome integrity and dynamics depends upon membrane tension and can be experimentally perturbed by osmotic swelling and deoxycholate treatment. We have also found that podosomes can be disrupted in a reversible manner by single or cyclic radial stretching of the substratum. We show that disruption of podosomes induced by osmotic swelling is independent of myosin-II filaments. Inhibition of the membrane sculpting protein, dynamin-II, but not clathrin, resulted in activation of myosin-IIA filament formation and disruption of podosomes. The effect of dynamin-II inhibition on podosomes was however independent of myosin-II filaments. Moreover, formation of organized arrays of podosomes in response to microtopographic cues (the ridges with triangular profile) was not accompanied by reorganization of myosin-II filaments. Thus, mechanical elements such as myosin-II filaments and factors affecting membrane tension/sculpting independently modulate podosome formation and dynamics, underlying a versatile response of these adhesion structures to intracellular and extracellular cues.

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Focal Adhesions Undergo Longitudinal Splitting into Fixed-Width Units

Cited by 33 publications

References 52 publications

Roles for Ena/VASP proteins in FMNL3-mediated filopodial assembly

Roles for Ena/VASP proteins in FMNL3-mediated filopodial assembly

Loss of Ena/VASP interferes with lamellipodium architecture, motility and integrin-dependent adhesion

Forces and constraints controlling podosome assembly and disassembly

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