Targeted disruption of vinculin genes in F9 and embryonic stem cells changes cell morphology, adhesion, and locomotion.

Abstract

Vinculin, a major constituent of focal adhesions and zonula adherens junctions, is thought to be involved in linking the microfilaments to areas of cell-substrate and cell-cell contacts. To test the role of vinculin in cell adhesion and motility, we used homologous recombination to generate F9 embryonal carcinoma and embryonic stem cell clones homozygous for a disrupted vinculin gene. When compared to wild-type cells, vinculin-mutant cells displayed a rounder morphology and a reduced ability to adhere and spread on plastic or fibronectin. Decreased adhesion of the mutant cells was associated with a reduction in lamellipodial extensions, as observed by time-lapse video microscopy. The locomotive activities of control F9 and the vinculin-null cells were compared in two assays. Loss of vinculin resulted in a 2.4-fold increase in cell motility. These results demonstrate an important role for vinculin in determining cell shape, adhesion, surface protrusive activity, and cell locomotion.

Expression of the focal adhesion protein paxillin in lung cancer and its relation to cell motility

“…The actin cytoskeleton interacts with the extracellular matrix and intracellular molecules via the focal adhesion. The focal adhesion contains proteins such as tensin and vinculin (Coll et al, 1995) which are thought to also have tumor suppressor properties. Focal adhesion proteins are also key regulators of some signal transduction events (Weisberg et al, 1997).…”

Intramolecular interactions regulate serine/threonine phosphorylation of vinculin

“…Although the precise biological function of vinculin is not fully understood yet, a variety of elegant studies has shown that vinculin is important in the organisation and maintenance of cell-contact architecture. Disruption of the vinculin gene leads to embryonic death in C. elegans [5] and to an altered adhesion and motility in F9 cells [6,7]. Similarly, cell adhesion is disturbed in cultered fibroblasts microinjected with antibodies against vinculin [8][9][10].…”

Temporal Dissection of β1-Integrin Signaling Indicates a Role for p130Cas-Crk in Filopodia Formation

“…Vinculin plays a central role in the mechanical coupling of integrins to the cytoskeleton as well as in related control of cytoskeletal mechanics, cell shape, and motility (13)(14)(15)(16). Talin is responsible for a conformational change of integrins to an active state, and it also links the receptor to the actin cytoskeleton (17)(18)(19)(20).…”