Force-initiated signal transduction can occur either via membrane-based ionic mechanisms or through changes in cytoskeletal–matrix linkages. We report here the stretch-dependent binding of cytoplasmic proteins to Triton X-100 cytoskeletons of L-929 cells grown on collagen-coated silicone. Triton X-100–insoluble cytoskeletons were stretched by 10% and incubated with biotinylated cytoplasmic proteins. Analysis with two-dimensional gel electrophoresis showed stretch-dependent binding of more than 10 cytoplasmic protein spots. Bound cytoplasmic proteins were purified by a photocleavable biotin tag and stretch-dependent binding of paxillin, focal adhesion kinase, and p130Cas was found, whereas the binding of vinculin was unchanged and actin binding decreased with stretch. Paxillin binding upon stretch was morphologically and biochemically similar in vitro and in vivo, that is, enhanced in the periphery and inhibited by the tyrosine phosphatase inhibitor, phenylarsine oxide. Thus, we suggest that transduction of matrix forces occurs through force-dependent conformation changes in the integrated cytoskeleton.
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18 February 2002
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February 11 2002
Force transduction by Triton cytoskeletons
Yasuhiro Sawada,
Yasuhiro Sawada
Department of Biological Sciences, Columbia University, New York, NY 10027
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Michael P. Sheetz
Michael P. Sheetz
Department of Biological Sciences, Columbia University, New York, NY 10027
Search for other works by this author on:
Yasuhiro Sawada
Department of Biological Sciences, Columbia University, New York, NY 10027
Michael P. Sheetz
Department of Biological Sciences, Columbia University, New York, NY 10027
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. E-mail: [email protected]
*
Abbreviations used in this paper: 2-D, two-dimensional; FAK, focal adhesion kinase; GFP, green fluorescent protein; PAO, phenylarsine oxide.
Received:
October 15 2001
Revision Received:
January 03 2002
Accepted:
January 03 2002
Online ISSN: 1540-8140
Print ISSN: 0021-9525
The Rockefeller University Press
2002
J Cell Biol (2002) 156 (4): 609–615.
Article history
Received:
October 15 2001
Revision Received:
January 03 2002
Accepted:
January 03 2002
Citation
Yasuhiro Sawada, Michael P. Sheetz; Force transduction by Triton cytoskeletons . J Cell Biol 18 February 2002; 156 (4): 609–615. doi: https://doi.org/10.1083/jcb.200110068
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