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Published online 9 January 2006. doi:10.1083/jcb.200506179
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 172, Number 2, 259-268
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Article

Focal adhesion size controls tension-dependent recruitment of {alpha}-smooth muscle actin to stress fibers

Jérôme M. Goffin1, Philippe Pittet1, Gabor Csucs2, Jost W. Lussi3, Jean-Jacques Meister1, and Boris Hinz1

1 Laboratory of Cell Biophysics, Ecole Polytechnique Fédérale de Lausanne, CH-1015, Lausanne, Switzerland
2 Institute of Biochemistry and 3 Institute for Biomedical Engineering, Swiss Federal Institute of Technology Zürich, CH-8093, Zürich, Switzerland

Correspondence to Boris Hinz: boris.hinz{at}epfl.ch

Expression of {alpha}-smooth muscle actin ({alpha}-SMA) renders fibroblasts highly contractile and hallmarks myofibroblast differentiation. We identify {alpha}-SMA as a mechanosensitive protein that is recruited to stress fibers under high tension. Generation of this threshold tension requires the anchoring of stress fibers at sites of 8–30-µm-long "supermature" focal adhesions (suFAs), which exert a stress approximately fourfold higher (~12 nN/µm2) on micropatterned deformable substrates than 2–6-µm-long classical FAs. Inhibition of suFA formation by growing myofibroblasts on substrates with a compliance of ≤11 kPa and on rigid micropatterns of 6-µm-long classical FA islets confines {alpha}-SMA to the cytosol. Reincorporation of {alpha}-SMA into stress fibers is established by stretching 6-µm-long classical FAs to 8.1-µm-long suFA islets on extendable membranes; the same stretch producing 5.4-µm-long classical FAs from initially 4-µm-long islets is without effect. We propose that the different molecular composition and higher phosphorylation of FAs on supermature islets, compared with FAs on classical islets, accounts for higher stress resistance.

Abbreviations used in this paper: AFM, atomic force microscopy; {alpha}-SMA, {alpha}-smooth muscle actin; FA, focal adhesion; FN, fibronectin; MHC, myosin heavy chain; MLC, myosin light chain; PDMS, polydimethylsiloxane; PLL-g-PEG, poly-L-lysine-g3.5-poly(ethylene glycol); REF, rat embryonic fibroblast; suFA, "supermature" FA; TX-100, Triton X-100; µCP, microcontact printing.


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