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© The Rockefeller University Press, 0021-9525/1998//1121 $5.00
The Journal of Cell Biology, Volume 142, Number 4, , 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 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 β1 integrin, although levels of 5 and 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 β1 integrin subunit were comparable to those in wild-type ES cells. We conclude that talin is essential for β1 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: talin • integrins • focal adhesions • gene knockout • ES cells

Abbreviations used in this paper: ANOVA, analysis of variance; ES cell, embryonic stem cell; Hyg, hygromycin; LIF, Leukemia inhibitory factor; Neo, neomycin; nt, nucleotides; TK, thymidine kinase.

Address all correspondence to D.R. Critchley, Department of Biochemistry, University of Leicester, University Road, Leicester LE1 7RH, UK. Tel.: 0116 252 3477. Fax: 0116 252 5097. E-mail: drc{at}leicester.ac.uk

Dr. Priddle's current address is Centre for Genome Research, University of Edinburgh, The King's Buildings, West Mains Rd., Edinburgh EH9 3JQ, UK.

Dr. Hemmings current address is Perkin Elmer, Applied Biosystems Division, 7 Kingsland Grange, Woolston, Warrington, Cheshire WA1 4SR, UK.

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