Integrin and receptor tyrosine kinase signalling networks cooperate to regulate various biological functions. The molecular details underlying the integration of both signalling networks remain largely uncharacterized. Here we identify a signalling module composed of a fibronectin–α5β1-integrin–integrin-linked-kinase (ILK) complex that, in concert with epidermal growth factor (EGF) cues, cooperatively controls the formation of transient actin-based circular dorsal ruffles (DRs) in fibroblasts. DR formation depends on the precise spatial activation of Src at focal adhesions by integrin and EGF receptor signals, in an ILK-dependent manner. In a SILAC-based phosphoproteomics screen we identified the tumour-suppressor Cyld as being required for DR formation induced by α5β1 integrin and EGF receptor co-signalling. Furthermore, EGF-induced Cyld tyrosine phosphorylation is controlled by integrin–ILK and Src as a prerequisite for DR formation. This study provides evidence for a novel function of integrin–ILK and EGF signalling crosstalk in mediating Cyld tyrosine phosphorylation and fast actin-based cytoskeletal rearrangements.
↵* Present address: The Beatson Institute for Cancer Research, Cancer Research UK Beatson Laboratories, Bearsden, Glasgow G611BD, UK
↵‡ Present address: Biomolecular Mass Spectrometry, Max Planck Institute for Heart and Lung Research, Bad Nauheim 61231, Germany
This work was supported by the Max Planck Society and the Tiroler Zukunftsstiftung.
Supplementary material available online at http://jcs.biologists.org/lookup/suppl/doi:10.1242/jcs.091652/-/DC1
- Accepted September 1, 2011.
- © 2012.