Conformation, localization, and integrin binding of talin depend on its interaction with phosphoinositides.
Talin is a structural component of focal adhesion sites and is thought to be engaged in multiple protein interactions at the cytoplasmic face of cell/matrix contacts. Talin is a major link between integrin and the actin cytoskeleton and was shown to play an important role in focal adhesion assembly. Consistent with the view that talin must be activated at these sites, we found that phosphatidylinositol 4-monophosphate and phosphatidylinositol 4,5-bisphosphate (PI4,5P(2)) bound to talin in cells in suspension or at early stages of adhesion, respectively. When phosphoinositides were associated with phospholipid bilayer, talin/phosphoinositide association was restricted to PI4,5P(2). This association led to a conformational change of the protein. Moreover, the interaction between integrin and talin was greatly enhanced by PI4,5P(2)-induced talin activation. Finally, sequestration of PI4,5P(2) by a specific pleckstrin homology domain confirms that PI4,5P(2) is necessary for proper membrane localization of talin and that this localization is essential for the maintenance of focal adhesions. Our results support a model in which PI4,5P(2) exposes the integrin-binding site on talin. We propose that PI4,5P(2)-dependent signaling modulates assembly of focal adhesions by regulating integrin-talin complexes. These results demonstrate that activation of the integrin-binding activity of talin requires not only integrin engagement to the extracellular matrix but also the binding of PI4,5P(2) to talin, suggesting a possible role of lipid metabolism in organizing the sequential assembly of focal adhesion components.
LEDAC, UMR CNRS/UJF 5538, Institut Albert Bonniot, Faculté de Médecine de Grenoble, 38706 La Tronche Cedex, France.
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V
Martel
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C
Racaud-Sultan
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Dupe
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Marie
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Paulhe
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Galmiche
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Block
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Albiges-Rizo
D016475:3T3 Cells; D000818:Animals; D001665:Binding Sites; D002448:Cell Adhesion / Q000502:physiology; D005353:Fibronectins / Q000502:physiology; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006801:Humans; D019012:Integrin beta1 / Q000502:physiology*; D007700:Kinetics; D008081:Liposomes; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics; D051379:Mice; D010716:Phosphatidylinositols / Q000378:metabolism* / Q000494:pharmacology; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D016608:Talin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D013917:Thrombin / Q000378:metabolism; D014162:Transfection
D005353:Fibronectins; D019012:Integrin beta1; D008081:Liposomes; D008164:Luminescent Proteins; D010716:Phosphatidylinositols; D011993:Recombinant Fusion Proteins; D016608:Talin; D049452:Green Fluorescent Proteins; D013917:Thrombin