Targeted disruption of vinculin genes in F9 and embryonic stem cells changes cell morphology, adhesion, and locomotion.
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.
Coll
J L
JL
La Jolla Cancer Research Foundation, CA 92037, USA.
Ben-Ze'ev
A
A
Ezzell
R M
RM
Rodríguez Fernández
J L
JL
Baribault
H
H
Oshima
R G
RG
Adamson
E D
ED
D000199:Actins / Q000032:analysis; D000818:Animals; D002448:Cell Adhesion; D002454:Cell Differentiation; D002460:Cell Line; D002465:Cell Movement; D002999:Clone Cells; D004622:Embryo, Mammalian; D004707:Endoderm; D005353:Fibronectins; D006801:Humans; D007700:Kinetics; D051379:Mice; D008969:Molecular Sequence Data; D015183:Restriction Mapping; D013234:Stem Cells / Q000166:cytology* / Q000502:physiology*; D018243:Teratocarcinoma; D014162:Transfection; D014407:Tumor Cells, Cultured; D016596:Vinculin / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics*
D000199:Actins; D005353:Fibronectins; D016596:Vinculin