Role of vinculin in regulating focal adhesion turnover

Saunders, Ruth; Holt, Mark R.; Jennings, Lisa K.; Sutton, Deborah H.; Barsukov, Igor; Bobkov, Andrey A.; Liddington, Robert; Adamson, Eileen A.; Dunn, Graham; Critchley, David R.

doi:10.1016/j.ejcb.2006.01.014

Cited by 167 publications

(195 citation statements)

References 51 publications

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“…1A). Consistent with previous studies [1][2][3][4], vinculin knockdown cells showed lower cell-cell and cell-ECM adhesions compared with mock-transfected HSC-4 cells (Fig. 1B).…”

Section: Vinculin Knockdown Promotes Mt1-mmp-mediated Prommp-2 Activasupporting

confidence: 92%

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Vinculin negatively regulates transcription of MT1-MMP through MEK/ERK pathway

Yoshimoto

Takino

et al. 2014

Biochemical and Biophysical Research Communications

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“…1A). Consistent with previous studies [1][2][3][4], vinculin knockdown cells showed lower cell-cell and cell-ECM adhesions compared with mock-transfected HSC-4 cells (Fig. 1B).…”

Section: Vinculin Knockdown Promotes Mt1-mmp-mediated Prommp-2 Activasupporting

confidence: 92%

“…Viculin has been implicated as an inhibitor of cell migration, as it enhanced cellular adhesiveness [1][2][3][4]. Loss or reduced expression of vinculin is related to the metastatic potential of squamous cell carcinomas (SCC), although in situ SCC exhibits high level expression of vinculin [5].…”

Section: Introductionmentioning

confidence: 99%

Vinculin negatively regulates transcription of MT1-MMP through MEK/ERK pathway

Yoshimoto

Takino

et al. 2014

Biochemical and Biophysical Research Communications

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“…1B), we further investigated the role of vinculin in FA architecture by mapping the positions of FA proteins in vinculin-null MEFs (32). As shown in Fig.…”

Section: Resultsmentioning

confidence: 99%

Talin determines the nanoscale architecture of focal adhesions

Liu

Wang

Goh

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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180

169

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Insight into how molecular machines perform their biological functions depends on knowledge of the spatial organization of the components, their connectivity, geometry, and organizational hierarchy. However, these parameters are difficult to determine in multicomponent assemblies such as integrin-based focal adhesions (FAs). We have previously applied 3D superresolution fluorescence microscopy to probe the spatial organization of major FA components, observing a nanoscale stratification of proteins between integrins and the actin cytoskeleton. Here we combine superresolution imaging techniques with a protein engineering approach to investigate how such nanoscale architecture arises. We demonstrate that talin plays a key structural role in regulating the nanoscale architecture of FAs, akin to a molecular ruler. Talin diagonally spans the FA core, with its N terminus at the membrane and C terminus demarcating the FA/stress fiber interface. In contrast, vinculin is found to be dispensable for specification of FA nanoscale architecture. Recombinant analogs of talin with modified lengths recapitulated its polarized orientation but altered the FA/stress fiber interface in a linear manner, consistent with its modular structure, and implicating the integrin-talin-actin complex as the primary mechanical linkage in FAs. Talin was found to be ∼97 nm in length and oriented at ∼15°relative to the plasma membrane. Our results identify talin as the primary determinant of FA nanoscale organization and suggest how multiple cellular forces may be integrated at adhesion sites.superresolution microscopy | focal adhesions | talin | mechanobiology | nanoscale architecture C ell adhesion to the ECM is a highly coordinated process that involves ECM-specific recognition by integrin transmembrane receptors, and their aggregation with numerous cytoplasmic proteins into dense supramolecular complexes called focal adhesions (FAs) (1). Actin stress fibers terminate at FAs where actomyosin contractility is transmitted to the ECM, generating traction (2-5). Mechanical tension impinging on each FA is implicated in key steps including the elongation, reinforcement, and maintenance of the FA structures (6). FA mechanotransduction is a major aspect of cellular microenvironment sensing with wide-ranging consequences in physiological and pathological processes (7-10). However, molecular-scale spatial parameters that specify FA nanoscale organization have been difficult to access experimentally. Nevertheless, these are essential to understand how mechanosensitivity arises within such complex molecular machines (11-15).Previously 3D superresolution fluorescence microscopy has unveiled the nanoscale organization of major FA components, whereby a core region of ∼30 nm interposes between the integrin and the actin cytoskeleton along the vertical (z) axis (16). The FA core consists of a membrane-proximal layer that contains signaling proteins such as FAK (focal adhesion kinase) and paxillin, an intermediate zone that contains force-transduction proteins s...

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“…The effect of vinculin on the migration of cells has previously been investigated using two-dimensional ECM substrates, where decreased vinculin expression caused increased cell migration (10). This finding has been explained by increased paxillin and focal adhesion kinase phosphorylation (11) and increased turnover of focal adhesions (12). In a recent study, we reported that vinculin acts as a mechano-regulating protein by increasing the generation of contractile forces (9).…”

mentioning

confidence: 97%

Vinculin Facilitates Cell Invasion into Three-dimensional Collagen Matrices

Mierke

Kollmannsberger

Zitterbart

et al. 2010

Journal of Biological Chemistry

180

175

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The cytoskeletal protein vinculin contributes to the mechanical link of the contractile actomyosin cytoskeleton to the extracellular matrix (ECM) through integrin receptors. In addition, vinculin modulates the dynamics of cell adhesions and is associated with decreased cell motility on two-dimensional ECM substrates. The effect of vinculin on cell invasion through dense three-dimensional ECM gels is unknown. Here, we report how vinculin expression affects cell invasion into three-dimensional collagen matrices. Cell motility was investigated in vinculin knockout and vinculin expressing wild-type mouse embryonic fibroblasts. Vinculin knockout cells were 2-fold more motile on two-dimensional collagen-coated substrates compared with wild-type cells, but 3-fold less invasive in 2.4 mg/ml three-dimensional collagen matrices. Vinculin knockout cells were softer and remodeled their cytoskeleton more dynamically, which is consistent with their enhanced two-dimensional motility but does not explain their reduced three-dimensional invasiveness. Importantly, vinculin-expressing cells adhered more strongly to collagen and generated 3-fold higher traction forces compared with vinculin knockout cells. Moreover, vinculin-expressing cells were able to migrate into dense (5.8 mg/ml) threedimensional collagen matrices that were impenetrable for vinculin knockout cells. These findings suggest that vinculin facilitates three-dimensional matrix invasion through up-regulation or enhanced transmission of traction forces that are needed to overcome the steric hindrance of ECMs.Cell migration is an important and fundamental biomechanical process that plays an essential role in inflammatory diseases, embryonic development, wound healing, and metastasis formation. Current concepts of cell migration have been established in two-dimensional models, but they can explain only partially the migratory behavior in three dimensions. For instance, the migratory capability of cells on two-dimensional substrates depends mainly on adhesion strength, adhesion dynamics, and the dynamics of cytoskeletal remodeling (1, 2), whereas the migratory capability of cells in three-dimensional connective tissue depends also on the steric hindrance of the matrix, matrix degradation by proteolytic enzyme secretion, and the generation of protrusive or contractile forces (1, 3-5). The balance of all these parameters-adhesion strength, cytoskeletal remodeling, matrix degradation, and the generation and transmission of contractile forces-is important for the migration speed in three-dimensional extracellular matrix (ECM) 2 (6). Depending on this balance, a broad variety of invasion strategies between different cell types and even within the same cell type are possible (7).The connection between the ECM and the actomyosin cytoskeleton through integrin-type cell-matrix adhesion receptors is facilitated by the mechano-coupling protein vinculin (8, 9). The effect of vinculin on the migration of cells has previously been investigated using two-dimensional ECM substrates, whe...

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Role of vinculin in regulating focal adhesion turnover

Cited by 167 publications

References 51 publications

Vinculin negatively regulates transcription of MT1-MMP through MEK/ERK pathway

Vinculin negatively regulates transcription of MT1-MMP through MEK/ERK pathway

Talin determines the nanoscale architecture of focal adhesions

Vinculin Facilitates Cell Invasion into Three-dimensional Collagen Matrices

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