Ezrin interacts with focal adhesion kinase and induces its activation independently of cell-matrix adhesion.
Ezrin, a membrane-cytoskeleton linker, is required for cell morphogenesis, motility, and survival through molecular mechanisms that remain to be elucidated. Using the N-terminal domain of ezrin as a bait, we found that p125 focal adhesion kinase (FAK) interacts with ezrin. We show that the two proteins coimmunoprecipitate from cultured cell lysates. However, FAK does not interact with full-length ezrin in vitro, indicating that the FAK binding site on ezrin is cryptic. Mapping experiments showed that the entire N-terminal domain of FAK (amino acids 1-376) is required for optimal ezrin binding. While investigating the role of the ezrin-FAK interaction, we observed that, in suspended kidney-derived epithelial LLC-PK1 cells, overproduction of ezrin promoted phosphorylation of FAK Tyr-397, the major autophosphorylation site, creating a docking site for FAK signaling partners. Treatment of the cells with a Src family kinase inhibitor reduced the phosphorylation of Tyr-577 but not that of Tyr-397, indicating that ezrin-mediated FAK activation does not require the activity of Src kinases. Altogether, these observations indicate that ezrin is able to trigger FAK activation in signaling events that are not elicited by cell-matrix adhesion.
Poullet
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P
Laboratoire de Morphogenèse et Signalisation Cellulaires, UMR 144 CNRS/Institut Curie, 26 rue d'Ulm, Paris 75248, cedex 05, France.
Gautreau
A
A
Kadaré
G
G
Girault
J A
JA
Louvard
D
D
Arpin
M
M
D000818:Animals; D021982:Cell-Matrix Junctions / Q000378:metabolism*; D003598:Cytoskeletal Proteins; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; D006801:Humans; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation; D011233:Precipitin Tests; D017434:Protein Structure, Tertiary; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D013552:Swine; D014407:Tumor Cells, Cultured; D014443:Tyrosine / Q000378:metabolism; D019061:src-Family Kinases / Q000378:metabolism; D000099149:Ezrin
D003598:Cytoskeletal Proteins; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; D010750:Phosphoproteins; D011505:Protein-Tyrosine Kinases; D014443:Tyrosine; D019061:src-Family Kinases; D000099149:Ezrin; C496050:PTK2 protein, human