Molecular shape and self‐association of vinculin and metavinculin

Molony, Leslie; Burridge, Keith

doi:10.1002/jcb.240290104

Cited by 87 publications

(56 citation statements)

References 14 publications

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“…They suggested that vinculin might selfassociate in focal contacts. Evidence for self-association of vinculin molecules has now been obtained from both direct binding assays (Otto, 1983;Belkin and Koteliansky, 1987) and EM (Milam, 1985;Molony and Burridge, 1985). We propose that self-association of vinculin molecules is responsible for the nonproportional inhibition of talin binding by unlabeled vinculin in our own experiments.…”

Section: Discussionsupporting

confidence: 58%

“…Thus, when a partial eDNA encoding just the COOH-terminal tail of vinculin is expressed in Cos cells, the polypeptide localizes to adhesion plaques, as does the vineulin encoded by cVin5 (Bendori et al, 1989). The tail region of vinculin is known to be a selfassociation site (Molony and Burridge, 1985;Milam, 1985), and it therefore seems likely that the expressed tail polypeptides are simply associating with endogenous monkey vineulin. The COOH-terminal tail region of vinculin is predicted to be basic relative to the globular head, which is predicted to be acidic (Coutu and Craig, 1988).…”

Section: Discussionmentioning

confidence: 99%

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Identification of a talin binding site in the cytoskeletal protein vinculin.

Jones

Jackson

Price

et al. 1989

The Journal of Cell Biology

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Abstract. Binding of the cytoskeletal protein vinculin to talin is one of a number of interactions involved in linking F-actin to cell-matrix junctions. To identify the talin binding domain in vinculin, we expressed the NH2-terminal region of the molecule encoded by two closely similar, but distinct vinculin cDNAs, using an in vitro transcription translation system. The 5' Eco RI-Bam HI fragment of a partial 2.89-kb vinculin cDNA encodes a 45-kD polypeptide containing the first 398 amino acids of the molecule. The equivalent restriction enzyme fragment of a second vinculin cDNA (cVin5) lacks nucleotides 746-867, and encodes a 41-kD polypeptide missing amino acids 167-207. The radiolabeled 45-kD vinculin polypeptide bound to microtiter wells coated with talin, but not BSA, and binding was inhibited by unlabeled vinculin. In contrast, the 41-kD vinculin polypeptide was devoid of talin binding activity. The role of residues 167-207 in talin binding was further analyzed by making a series of deletions spanning this region, each deletion of seven amino acids contiguous with the next.

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Section: Discussionsupporting

confidence: 58%

Section: Discussionmentioning

confidence: 99%

Identification of a talin binding site in the cytoskeletal protein vinculin.

Jones

Jackson

Price

et al. 1989

The Journal of Cell Biology

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“…As shown in supplemental Fig. S3C, multiple higher order Vt species form in the absence of actin and the ability for vinculin to form these higher order oligomers has been previously observed (46,47). Because these higher order Vt oligomers disappear with increasing actin concentrations and because Vt ⌬C5 is a F-actin bundling-deficient mutant, we have chosen to focus on changes in the dimer and trimer bands between WT Vt and Vt ⌬C5 with increasing concentrations of actin.…”

Section: Vt Association With F-actin Promotes Vt Dimerization-vtsupporting

confidence: 65%

The Vinculin C-terminal Hairpin Mediates F-actin Bundle Formation, Focal Adhesion, and Cell Mechanical Properties

Shen

Tolbert

Guilluy

et al. 2011

Journal of Biological Chemistry

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Vinculin is an essential and highly conserved cell adhesion protein, found at both focal adhesions and adherens junctions, where it couples integrins or cadherins to the actin cytoskeleton. Vinculin is involved in controlling cell shape, motility, and cell survival, and has more recently been shown to play a role in force transduction. The tail domain of vinculin (Vt) contains determinants necessary for binding and bundling of actin filaments. Actin binding to Vt has been proposed to induce formation of a Vt dimer that is necessary for cross-linking actin filaments. Results from this study provide additional support for actininduced Vt self-association. Moreover, the actin-induced Vt dimer appears distinct from the dimer formed in the absence of actin. To better characterize the role of the Vt strap and carboxyl terminus (CT) in actin binding, Vt self-association, and actin bundling, we employed smaller amino-terminal (NT) and CT deletions that do not perturb the structural integrity of Vt. Although both NT and CT deletions retain actin binding, removal of the CT hairpin (1061-1066) selectively impairs actin bundling in vitro. Moreover, expression of vinculin lacking the CT hairpin in vinculin knock-out murine embryonic fibroblasts affects the number of focal adhesions formed, cell spreading as well as cellular stiffening in response to mechanical force.The ability of cells to sense and respond to environmental cues such as mechanical forces is critical for multiple cellular processes including embryogenesis and wound healing (1-3). To transmit forces across the cell membrane in both directions, the actin cytoskeleton couples transmembrane receptors (integrin or cadherin), through points of cell adhesion consisting of multiple protein complexes that form cell-extracellular matrix (focal adhesions) and cell-cell (adherens junctions) contacts (3, 4). Vinculin is an abundant protein found in both focal adhesions and adherens junctions, and plays a key role in regulating cell morphology, cell motility, and force transduction (2, 5). Vinculin is essential during development as vinculin knockout (KO) mouse embryos fail to survive beyond day E10 with extensive defects in myocardial and endocardial structures (6). Consistent with a role in muscle structure, vinculin KO mice are predisposed to stress-induced cardiomyopathy (7). Moreover, mutations and deletions in the tail domain of metavinculin, a splice isoform of vinculin, are associated with dilated cardiomyopathy (8 -10). Vinculin also possesses tumor suppressor properties as vinculin KO cells are less adherent, have a rounded morphology, reduced lamellipodial stability, increased motility (6, 11), and are resistant to apoptosis and anoikis (12).Vinculin is a highly conserved cytoskeletal protein that has 1066 residues and contains a globular head, a flexible prolinerich linker, and a tail domain (13). Each discrete vinculin domain recognizes multiple binding partners: the vinculin head (Vh) 4 binds to talin, ␣-actinin, and IpaA (14 -16); the prolinerich linker inte...

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“…This study demonstrates, however, that vinculin binds to E-cadherin complexes in the absence of ␣-catenin, which eliminates binding to ␣-catenin as the sole mechanism for the targeting of vinculin to the adhesion complex. The possibility that vinculin may cross-link adhesion complexes is inferred from electron microscopy studies of rotary-shadowed vinculin that demonstrate the existence in vitro of vinculin complexes containing two to six individual molecules (45,46). It remains to be determined whether this multimerization takes place in vivo.…”

Section: Discussionmentioning

confidence: 99%

Vinculin Is Associated with the E-cadherin Adhesion Complex

Hazan¹,

Kang²,

Roe³

et al. 1997

Journal of Biological Chemistry

156

130

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Cadherins mediate calcium-dependent cell-cell adhesion, and this activity is regulated by cytoplasmic interactions between cadherins, catenins, and the actinbased cytoskeleton. ␣-Catenin plays a critical role in the transmembrane anchorage of cadherins, and deletion of ␣-catenin has been shown to inactivate cadherin-mediated adhesion, resulting in a nonadhesive phenotype. Here we show that serum starvation increases E-cadherin expression and induces E-cadherin-dependent adhesion in the MDA-MB-468 breast cancer cell line. This adhesion occurred despite a lack of ␣-catenin expression, which was caused by mutations in the ␣-catenin gene. Coprecipitation analysis suggests that this adhesion may be mediated by cytoplasmic connections from cadherins to the cytoskeleton involving vinculin. A high level of vinculin associated with E-cadherin immunoprecipitates was observed in MDA-MB-468 cells. In contrast, vinculin was not detected in E-cadherin complexes in the A431 and MCF-7 epithelial carcinoma cell lines, which express ␣-catenin. However, in reciprocal immunoprecipitations using anti-vinculin antibodies, Ecadherin associated strongly with vinculin in MDA-MB-468 cells and, to a lesser extent, in A431 and MCF-7 cells. These results suggest that both ␣-catenin and vinculin may be present in the adhesion complex. To test the hypothesis that vinculin associates with E-cadherin complexes via ␤-catenin, excess recombinant ␤-catenin or ␣-catenin fusion protein was added to MDA-MB-468 cell lysates. Both specifically inhibited the coprecipitation of E-cadherin with vinculin, suggesting competition for the same binding site. These results suggest that vinculin plays a role in the establishment or regulation of the cadherin-based cell adhesion complex by direct interaction with ␤-catenin.Cadherins are calcium-dependent cell-cell adhesion molecules that mediate homotypic interactions among cells and are essential for tissue morphogenesis (for review, see Refs. 1 and 2). Adhesion via cadherins involves the coordination of extracellular binding and intracellular anchorage to the actin-based cytoskeleton. The cytoplasmic domain of E-cadherin binds to either ␤-catenin or plakoglobin/␥-catenin (for review, see , and this complex is coupled to the actin cytoskeleton by ␣-catenin, which binds to both ␤-catenin and actin (6, 7). There is evidence that ␣-catenin also binds to ␣-actinin (8, 9) and spectrin (10), but the role of these interactions in adhesion is unknown. Disruption of ␣-catenin function by genetic deletion or mutation was shown to cause a loss of E-cadherin-dependent adhesion. PC-9 lung carcinoma cells, which lack detectable ␣-catenin expression, were shown to have aberrant cell-cell adhesion (11) that was restored by transfection of these cells with ␣-catenin cDNA (12). This resulted in the establishment of a polarized epithelium and inhibition of cell growth (13). This and other work suggest that the linkage of cadherin complexes to the cytoskeleton via ␣-catenin is essential for normal cell adhesion, morphogenesis, and ...

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Molecular shape and self‐association of vinculin and metavinculin

Cited by 87 publications

References 14 publications

Identification of a talin binding site in the cytoskeletal protein vinculin.

Identification of a talin binding site in the cytoskeletal protein vinculin.

The Vinculin C-terminal Hairpin Mediates F-actin Bundle Formation, Focal Adhesion, and Cell Mechanical Properties

Vinculin Is Associated with the E-cadherin Adhesion Complex

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