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This study for the first time reports the specific binding of the alternatively spliced EIIIA (ED-A) segment of FN to integrins α9β1 and α4β1, of which the latter is expressed in fibroblastic cells. ED-A FN has previously been shown to be crucial for myofibroblast differentiation [9] and binding of α4β1 integrin appears to be crucial for stress-activation of ERK2 [70]. Thus, identification of α4β1 integrin as the specific ED-A FN receptor is an important step to decipher the mechanisms of ECM stress-induced expression of α-SMA
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Liao Y.F., Gotwals P.J., Koteliansky V.E., Sheppard D., Van De Water L. The EIIIA segment of fibronectin is a ligand for integrins α9β1 and α4β1 providing a novel mechanism for regulating cell adhesion by alternative splicing. J. Biol. Chem. 277:2002;14467-14474 This study for the first time reports the specific binding of the alternatively spliced EIIIA (ED-A) segment of FN to integrins α9β1 and α4β1, of which the latter is expressed in fibroblastic cells. ED-A FN has previously been shown to be crucial for myofibroblast differentiation [9] and binding of α4β1 integrin appears to be crucial for stress-activation of ERK2 [70]. Thus, identification of α4β1 integrin as the specific ED-A FN receptor is an important step to decipher the mechanisms of ECM stress-induced expression of α-SMA.
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48
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Patel K., Harding P., Haney L.B., Glass W.F. II Regulation of the mesangial cell myofibroblast phenotype by actin polymerization. J. Cell Physiol. 195:2003;435-445.
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In an elegant approach, the authors subjected Triton-insoluble cytoskeletons to mechanical stress using silicone membranes and incubated them with extracted cytoplasmic proteins. The major finding of this work is that FA proteins paxillin and FAK, but not vinculin and actin, selectively incorporate into mechanically stressed Triton-cytoskeletons. This finding suggests that a stress-dependent conformational change in the cytoskeleton creates binding sites (or space) for the incorporation of cytoplasmic proteins at sites of FAs.
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Sawada Y., Sheetz M.P. Force transduction by Triton cytoskeletons. J. Cell Biol. 156:2002;609-615 In an elegant approach, the authors subjected Triton-insoluble cytoskeletons to mechanical stress using silicone membranes and incubated them with extracted cytoplasmic proteins. The major finding of this work is that FA proteins paxillin and FAK, but not vinculin and actin, selectively incorporate into mechanically stressed Triton-cytoskeletons. This finding suggests that a stress-dependent conformational change in the cytoskeleton creates binding sites (or space) for the incorporation of cytoplasmic proteins at sites of FAs.
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Riveline D., Zamir E., Balaban N.Q., Schwarz U.S., Ishizaki T., Narumiya S., Kam Z., Geiger B., Bershadsky A.D. Focal contacts as mechanosensors: externally applied local mechanical force induces growth of focal contacts by an mDia1-dependent and ROCK-independent mechanism. J. Cell Biol. 153:2001;1175-1186.
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Cells lying on a bed of microneedles: An approach to isolate mechanical force
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This work makes significant progress in the attempt to measure forces on the level of single FAs by combining the use of elastic polymer substrates, pioneered by Harris et al. [26] and by Pelham and Wang [28], and of microcontact printed surfaces, as originally developed by Ingber [30]. Cells are grown on an array of microposts with a cross-sectional area in the range of the size of FAs; deflection of the posts provides a direct measure for the intracellular contractile forces transmitted by individual FAs and is not influenced by the force developed at FAs on neighboring posts. Using this technique, the authors demonstrated similar size/force ratio of FAs compared with the work of Geiger and co-workers (reviewed in [71]) without requiring extensive mathematical modeling. Moreover, it allows changing substrate compliance by adjusting the height of the posts (and thus their deformability) by keeping the chemical properties of the polymer constant
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Tan J.L., Tien J., Pirone D.M., Gray D.S., Bhadriraju K., Chen C.S. Cells lying on a bed of microneedles: an approach to isolate mechanical force. Proc. Natl. Acad. Sci. U.S.A. 100:2003;1484-1489 This work makes significant progress in the attempt to measure forces on the level of single FAs by combining the use of elastic polymer substrates, pioneered by Harris et al. [26] and by Pelham and Wang [28], and of microcontact printed surfaces, as originally developed by Ingber [30]. Cells are grown on an array of microposts with a cross-sectional area in the range of the size of FAs; deflection of the posts provides a direct measure for the intracellular contractile forces transmitted by individual FAs and is not influenced by the force developed at FAs on neighboring posts. Using this technique, the authors demonstrated similar size/force ratio of FAs compared with the work of Geiger and co-workers (reviewed in [71]) without requiring extensive mathematical modeling. Moreover, it allows changing substrate compliance by adjusting the height of the posts (and thus their deformability) by keeping the chemical properties of the polymer constant.
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