Disruption of the talin gene compromises focal adhesion assembly in undifferentiated but not differentiated embryonic stem cells.
We have used gene disruption to isolate two talin (-/-) ES cell mutants that contain no intact talin. The undifferentiated cells (a) were unable to spread on gelatin or laminin and grew as rounded colonies, although they were able to spread on fibronectin (b) showed reduced adhesion to laminin, but not fibronectin (c) expressed much reduced levels of beta1 integrin, although levels of alpha5 and alphaV were wild-type (d) were less polarized with increased membrane protrusions compared with a vinculin (-/-) ES cell mutant (e) were unable to assemble vinculin or paxillin-containing focal adhesions or actin stress fibers on fibronectin, whereas vinculin (-/-) ES cells were able to assemble talin-containing focal adhesions. Both talin (-/-) ES cell mutants formed embryoid bodies, but differentiation was restricted to two morphologically distinct cell types. Interestingly, these differentiated talin (-/-) ES cells were able to spread and form focal adhesion-like structures containing vinculin and paxillin on fibronectin. Moreover, the levels of the beta1 integrin subunit were comparable to those in wild-type ES cells. We conclude that talin is essential for beta1 integrin expression and focal adhesion assembly in undifferentiated ES cells, but that a subset of differentiated cells are talin independent for both characteristics.
Priddle
H
H
Department of Biochemistry, University of Leicester, Leicester LE1 7RH, United Kingdom.
Hemmings
L
L
Monkley
S
S
Woods
A
A
Patel
B
B
Sutton
D
D
Dunn
G A
GA
Zicha
D
D
Critchley
D R
DR
D000199:Actins / Q000378:metabolism; D000818:Animals; D002448:Cell Adhesion / Q000502:physiology*; D002454:Cell Differentiation / Q000502:physiology*; D002455:Cell Division / Q000235:genetics; D002934:Cinnamates; D003598:Cytoskeletal Proteins / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D005780:Gelatin / Q000378:metabolism; D018390:Gene Targeting / Q000379:methods; D006921:Hygromycin B / Q000031:analogs & derivatives / Q000378:metabolism; D016023:Integrins / Q000378:metabolism; D007797:Laminin / Q000378:metabolism; D051379:Mice; D009154:Mutation / Q000235:genetics; D051419:Paxillin; D010641:Phenotype; D010750:Phosphoproteins / Q000378:metabolism; D013234:Stem Cells / Q000502:physiology*; D016608:Talin / Q000172:deficiency* / Q000235:genetics; D016596:Vinculin / Q000235:genetics
D000199:Actins; D002934:Cinnamates; D003598:Cytoskeletal Proteins; D005353:Fibronectins; D016023:Integrins; D007797:Laminin; D051419:Paxillin; D010750:Phosphoproteins; C496062:Pxn protein, mouse; D016608:Talin; D016596:Vinculin; D006921:Hygromycin B; C026273:hygromycin A; D005780:Gelatin