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Published 27 October 2003. doi:10.1083/jcb.200302001
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© The Rockefeller University Press, 0021-9525/2003/10/409 $8.00
The Journal of Cell Biology, Volume 163, Number 2, 409-419

Article

 Talin1 is critical for force-dependent reinforcement of initial integrin–cytoskeleton bonds but not tyrosine kinase activation

Grégory Giannone1, Guoying Jiang1, Deborah H. Sutton2, David R. Critchley2 and Michael P. Sheetz1

1 Department of Biological Sciences, Columbia University, New York, NY 10027
2 Department of Biochemistry, University of Leicester, Leicester LE1 7RH, UK

Address correspondence to Michael P. Sheetz, Dept. of Biological Sciences, P.O. Box 2408, Columbia University, Sherman Fairchild Center, Rm. 713, 1212 Amsterdam Ave., New York, NY 10027. Tel.: (212) 854-4857. Fax: (212) 854-6399. email: ms2001{at}columbia.edu

Cells rapidly transduce forces exerted on extracellular matrix contacts into tyrosine kinase activation and recruitment of cytoskeletal proteins to reinforce integrin–cytoskeleton connections and initiate adhesion site formation. The relationship between these two processes has not been defined, particularly at the submicrometer level. Using talin1-deficient cells, it appears that talin1 is critical for building early mechanical linkages. Deletion of talin1 blocked laser tweezers, force-dependent reinforcement of submicrometer fibronectin-coated beads and early formation of adhesion sites in response to force, even though Src family kinases, focal adhesion kinase, and spreading were activated normally. Recruitment of vinculin and paxillin to sites of force application also required talin1. FilaminA had a secondary role in strengthening fibronectin–integrin–cytoskeleton connections and no role in stretch-dependent adhesion site assembly. Thus, force-dependent activation of tyrosine kinases is independent of early force-dependent structural changes that require talin1 as part of a critical scaffold.

Key Words: talin; integrin; actin cytoskeleton; tyrosine phosphorylation; mechano-sensing

Abbreviations used in this paper: ABS, actin-binding site; ES, embryonic stem; FN, fibronectin; MSD, mean square displacement; RPTP{alpha}, receptor-like protein phosphatase {alpha}; SFK, Src family kinase; TIRF, total internal reflection fluorescence; VN, vitronectin.


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