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J. Cell Biol., Volume 142, Number 4, August 24, 1998 1121-1133

Disruption of the Talin Gene Compromises Focal Adhesion Assembly in Undifferentiated but Not Differentiated Embryonic Stem Cells

Helen Priddle,* Lance Hemmings,* Susan Monkley,* Alison Woods,* Bipin Patel,* Deborah Sutton,* Graham A. Dunn,Dagger Daniel Zicha,Dagger and David R. Critchley*

* Department of Biochemistry, University of Leicester, Leicester LE1 7RH, United Kingdom; and Dagger  Medical Research Council Muscle and Cell Motility Unit, The Randall Institute, King's College London, 26-29 Drury Lane, London WC2B 5RL, United Kingdom

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 beta 1 integrin, although levels of alpha 5 and alpha V 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 beta 1 integrin subunit were comparable to those in wild-type ES cells. We conclude that talin is essential for beta 1 integrin expression and focal adhesion assembly in undifferentiated ES cells, but that a subset of differentiated cells are talin independent for both characteristics.

Key words: talinintegrinsfocal adhesionsgene knockoutES cells


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