Vinculin regulates directionality and cell polarity in two- and three-dimensional matrix and three-dimensional microtrack migration

Rahman, Aniqua; Carey, Shawn P.; Kraning-Rush, Casey M.; Goldblatt, Zachary E.; Bordeleau, François; Lampi, Marsha C.; Lin, Deanna; Garcı́a, Andrés J.; Reinhart‐King, Cynthia A.

doi:10.1091/mbc.e15-06-0432

Cited by 59 publications

(73 citation statements)

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“…Thus, the directional asymmetry of the vinculin-actin bond is sufficient to establish a spatial asymmetry in F-actin polarity (Fig. S7), an effect that may underlie the prior observation that vinculin is essential for generating persistent, directional cell migration (12,13,30). The directionally asymmetric linkage of vinculin to actin may complement other mechanisms by which cells generate persistent migration, such as the signaling cascade that activates the barbed (+) end nucleators Arp2/3 (31) and formins (32), which reinforce cell polarity by promoting actin polymerization at the cell's leading edge.…”

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confidence: 68%

“…Vinculin plays a key role in maintaining tissue integrity (9,10); for example, loss of vinculin in mice results in the death of the developing embryo owing to defects in neural tube closure and heart development (11). Importantly, vinculin is required for persistent directional cell migration, suggestive of a role in generating a polarized connection between adhesions and the actin cytoskeleton (12,13). Although vinculin is also recruited to cadherin-based adhesions in a force-dependent manner (6,14), comparatively little is known about how it might regulate actin organization and dynamics at those sites.…”

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confidence: 99%

“…Consistent with this idea, vinculin null mouse embryos exhibit phenotypes suggestive of failures not only in cell adhesion, but also in tissue patterning driven by cell migration, notably axon outgrowth (11). Vinculin-deficient cells likewise exhibit defects in the formation of stable lamellipodia and filopodia (36), and are more motile (37) but less directionally persistent (13,30). Thus, vinculin does not appear to be required for adhesion or motility per se, but instead, for stable protrusion and polarized migration.…”

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Vinculin Forms a Directionally Asymmetric Catch Bond with F-Actin

Huang

Bax

Buckley

et al. 2018

Biophysical Journal

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Vinculin is an actin-binding protein thought to reinforce cell-cell and cell-matrix adhesions. However, how mechanical load affects the vinculin/F-actin bond is unclear. Using a singlemolecule optical trap assay, we found that vinculin forms a force-dependent catch bond with Factin via its tail domain, but with lifetimes that depend strongly on the direction of the applied force. Force toward the pointed end of the actin filament resulted in a bond that was maximally stable at 8 pN, with a mean lifetime (12 s) 10-fold longer than the mean lifetime when force was applied toward the barbed end. A computational model of lamellipodial actin dynamics suggested that the directionality of the vinculin/F-actin bond could establish long-range order in the actin cytoskeleton. The directional and force-stabilized binding of vinculin to F-actin may provide a mechanism by which adhesion complexes maintain front-rear asymmetry in migrating cells.Cadherin-and integrin-based protein assemblies link cells to each other and to the extracellular matrix (ECM), respectively, and together provide the physical basis for the organization of multicellular tissues (1). Both classes of adhesion complexes are exquisitely sensitive to mechanical load, and change rapidly in size and composition in order to maintain the physical integrity of living tissues (2). These adhesions are also essential in defining the physical asymmetries that underlie both individual and collective cell migration in the context of embryonic development (3), wound healing (4), and cancer metastasis (5). However, the molecular basis of how cadherin-and integrin-based adhesions respond to The protein vinculin is a component of both cadherin-and integrin-based adhesion complexes, and is rapidly recruited to both types of adhesions in response to mechanical load through its interactions with α-catenin and talin, respectively (6-8). Vinculin plays a key role in maintaining tissue integrity (9, 10); for example, loss of vinculin in mice results in the death of the developing embryo owing to defects in neural tube closure and heart development (11). Importantly, vinculin is required for persistent directional cell migration, suggestive of a role in generating a polarized connection between adhesions and the actin cytoskeleton (12, 13). Although vinculin is also recruited to cadherin-based adhesions in a force-dependent manner (6, 14), comparatively little is known about how it might regulate actin organization and dynamics at those sites.Vinculin binds directly to filamentous (F-) actin through its actin-binding tail domain (Vt) (15, 16), but how and whether this bond may be regulated by mechanical load is not known. Defining this mechanism is critical to understanding the role of vinculin as a reinforcing link between adhesion complexes and the actin cytoskeleton. We modified a previously developed optical trap (OT)-based assay (17) to define the load dependence of the binding interaction between vinculin and F-actin (Fig. 1A). Actin binding to full-length, wild-t...

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Vinculin Forms a Directionally Asymmetric Catch Bond with F-Actin

Huang

Bax

Buckley

et al. 2018

Biophysical Journal

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“…Vinculin plays a key role in maintaining tissue integrity (9, 10); for example, loss of vinculin in mice results in the death of the developing embryo owing to defects in neural tube closure and heart development (11). Importantly, vinculin is required for persistent directional cell migration, suggestive of a role in generating a polarized connection between adhesions and the actin cytoskeleton (12, 13). Although vinculin is also recruited to cadherin-based adhesions in a force-dependent manner (6, 14), comparatively little is known about how it might regulate actin organization and dynamics at those sites.…”

mentioning

confidence: 99%

Vinculin forms a directionally asymmetric catch bond with F-actin

Huang

Bax

Buckley

et al. 2017

Science

235

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“…During this process, vinculin which has over 14 binding partners, has to be activated from the auto-inhibitory self-binding state. Vinculin regulates directionality and cell polarity, and the knockdown showed reduced traction force and polarity loss [35]. As shown in Fig 11, well-developed actin stress fibers were confirmed independent of αB-crystallin expression but position of matured focal adhesion as visualized by vinculin immuno-staining, stress fiber direction, length, and density were clearly αB-crystallin dependent.…”

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Small Heat Shock Protein αB-Crystallin Controls Shape and Adhesion of Glioma and Myoblast Cells in the Absence of Stress

2016

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Cell shape and adhesion and their proper controls are fundamental for all biological systems. Mesenchymal cells migrate at an average rate of 6 to 60 μm/hr, depending on the extracellular matrix environment and cell signaling. Myotubes, fully differentiated muscle cells, are specialized for power-generation and therefore lose motility. Cell spreading and stabilities of focal adhesion are regulated by the critical protein vinculin from immature myoblast to mature costamere of differentiated myotubes where myofibril Z-band linked to sarcolemma. The Z-band is constituted from microtubules, intermediate filaments, cell adhesion molecules and other adapter proteins that communicate with the outer environment. Mesenchymal cells, including myoblast cells, convert actomyosin contraction forces to tension through mechano-responsive adhesion assembly complexes as Z-band equivalents. There is growing evidence that microtubule dynamics are involved in the generation of contractile forces; however, the roles of microtubules in cell adhesion dynamics are not well determined. Here, we show for the first time that αB-crystallin, a molecular chaperon for tubulin/microtubules, is involved in cell shape determination. Moreover, knockdown of this molecule caused myoblasts and glioma cells to lose their ability for adhesion as they tended to behave like migratory cells. Surprisingly, αB-crystallin knockdown in both C6 glial cells and L6 myoblast permitted cells to migrate more rapidly (2.7 times faster for C6 and 1.3 times faster for L6 cells) than dermal fibroblast. On the other hand, overexpression of αB-crystallin in cells led to an immortal phenotype because of persistent adhesion. Position of matured focal adhesion as visualized by vinculin immuno-staining, stress fiber direction, length, and density were clearly αB-crystallin dependent. These results indicate that the small HSP αB-crystallin has important roles for cell adhesion, and thus microtubule dynamics are necessary for persistent adhesion.

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Vinculin regulates directionality and cell polarity in two- and three-dimensional matrix and three-dimensional microtrack migration

Cited by 59 publications

References 50 publications

Vinculin Forms a Directionally Asymmetric Catch Bond with F-Actin

Vinculin Forms a Directionally Asymmetric Catch Bond with F-Actin

Vinculin forms a directionally asymmetric catch bond with F-actin

Small Heat Shock Protein αB-Crystallin Controls Shape and Adhesion of Glioma and Myoblast Cells in the Absence of Stress

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