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. 2008 Dec 10;66(2):324–338. doi: 10.1007/s00018-008-8424-4

Src activation and translocation from focal adhesions to membrane ruffles contribute to formation of new adhesion sites

A Hamadi 1, T B Deramaudt 1, K Takeda 1, P Rondé 1,
PMCID: PMC11131495  PMID: 19066724

Abstract.

Cell migration requires the coordinated turnover of focal adhesions, a process that involves FAK phosphorylation. Since Src is the major kinase implicated in FAK phosphorylation, we focus here on the role of Src activation on adhesion remodelling. In astrocytoma cells, constitutively activated Src induces both FAK phosphorylation and adhesion rearrangement. To evaluate how Src controls these processes, we used a recently described Src reporter to monitor the dynamics of Src phosphorylation. Upon Src activation, focal adhesions started to disassemble while Src appeared highly expressed at newly formed membrane ruffles. Kinetic analysis of time-lapse movies showed that loss of phospho-Src at focal adhesions was time-correlated with the appearance of membrane ruffles containing phospho-Src. Moreover, FLIP analysis revealed a dynamic equilibrium of Src between focal adhesions and membrane ruffles. We conclude that upon phosphorylation, Src is directly translocated from focal adhesions to membrane ruffles, thereby promoting formation of new adhesion complexes.

Electronic supplementary material

The online version of this article (doi:10.1007/s00018-008-8424-4) contains supplementary material, which is available to authorized users.

Keywords. Src, tyrosine phosphorylation, Src indicator, focal adhesion dynamics, FAK

Electronic supplementary material

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ESM1 (MPG 57 kb) (56.7KB, mpg)
ESM2 (MPG 1.60 MB) (1.6MB, mpg)
ESM3 (MPG 253 kb) (253.4KB, mpg)
ESM3 (MPG 160 kb) (160.6KB, mpg)
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Footnotes

Received 21 July 2008; received after revision 10 October 2008; accepted 03 November 2008

Associated Data

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Supplementary Materials

ESM1 (MPG 57 kb) (56.7KB, mpg)
ESM2 (MPG 1.60 MB) (1.6MB, mpg)
ESM3 (MPG 253 kb) (253.4KB, mpg)
ESM3 (MPG 160 kb) (160.6KB, mpg)
ESM3 (MPG 743 kb) (743.2KB, mpg)
(PDF 52 kb) (51.6KB, pdf)

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