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

doi:10.1083/jcb.142.4.1121

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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, Daniel Zicha, and David R. Critchley^*

^* Department of Biochemistry, University of Leicester, Leicester LE1 7RH, United Kingdom; and 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 undifferentiatedcells (a) were unable to spread on gelatin or laminin and grewas 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 levelsof $alpha$ 5 and $alpha$ V were wild-type (d) were less polarized with increasedmembrane protrusions compared with a vinculin ( $-$ / $-$ ) ES cell mutant(e) were unable to assemble vinculin or paxillin-containing focaladhesions 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 differentiationwas restricted to two morphologically distinct cell types. Interestingly,these differentiated talin ( $-$ / $-$ ) ES cells were able to spreadand form focal adhesion-like structures containing vinculin andpaxillin on fibronectin. Moreover, the levels of the $beta$ 1 integrinsubunit were comparable to those in wild-type ES cells. We concludethat talin is essential for $beta$ 1 integrin expression and focal adhesionassembly in undifferentiated ES cells, but that a subset of differentiatedcells are talin independent for both characteristics.

Key words: talin; integrins; focal adhesions; gene knockout; ES cells

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