Abstract
Cell migration is a complex, highly regulated process that involves the continuous formation and disassembly of adhesions (adhesion turnover). Adhesion formation takes place at the leading edge of protrusions, whereas disassembly occurs both at the cell rear and at the base of protrusions. Despite the importance of these processes in migration, the mechanisms that regulate adhesion formation and disassembly remain largely unknown. Here we develop quantitative assays to measure the rate of incorporation of molecules into adhesions and the departure of these proteins from adhesions. Using these assays, we show that kinases and adaptor molecules, including focal adhesion kinase (FAK), Src, p130CAS, paxillin, extracellular signal-regulated kinase (ERK) and myosin light-chain kinase (MLCK) are critical for adhesion turnover at the cell front, a process central to migration.
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Acknowledgements
We thank M. Beckerle, K. Rottner, A. Bouton, A. Hall, J. Collard, S. Parsons, M. Weber and D. Ilic for reagents and suggestions. This work was supported by NIH grants GM23244 (A.F.H.), CA40042 (J.T.P.) and GM47607 (C.E.T.) and by the University of Virginia Cancer Center. D.J.W. was supported by NIH postdoctoral training grant HD07528-01.
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Supplementary Fig. 1 (PDF 170 kb)
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Supplementary Movie S1
Time-lapse images for Fig. 1 showing an MEF expressing paxillin-GFP. (MOV 130 kb)
Supplementary Movie S2
Time-lapse images for Fig. 1, which shows an MEF expressing paxillin-DsRed2. (MOV 149 kb)
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Webb, D., Donais, K., Whitmore, L. et al. FAK–Src signalling through paxillin, ERK and MLCK regulates adhesion disassembly. Nat Cell Biol 6, 154–161 (2004). https://doi.org/10.1038/ncb1094
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DOI: https://doi.org/10.1038/ncb1094
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