title: Phosphorylation of N-cadherin-associated cortactin by Fer kinase regulates N-cadherin mobility and intercellular adhesion strength.
abstract: Cortactin regulates the strength of nascent N-cadherin-mediated intercellular adhesions through a tyrosine phosphorylation-dependent mechanism. Currently, the functional significance of cortactin phosphorylation and the kinases responsible for the regulation of adhesion strength are not defined. We show that the nonreceptor tyrosine kinase Fer phosphorylates cadherin-associated cortactin and that this process is involved in mediating intercellular adhesion strength. In wild-type fibroblasts N-cadherin ligation-induced transient phosphorylation of Fer, indicating that junction formation activates Fer kinase. Tyrosine phosphorylation of cortactin after N-cadherin ligation was strongly reduced in fibroblasts expressing only catalytically inactive Fer (D743R), compared with wild-type cells. In wild-type cells, N-cadherin-coated bead pull-off assays induced fourfold greater endogenous N-cadherin association than in D743R cells. Fluorescence recovery after photobleaching showed that GFP-N-cadherin mobility at nascent contacts was 50% faster in wild-type than D743R cells. In shear wash-off assays, nascent intercellular adhesion strength was twofold higher in wild-type than D743R cells. Cortactin recruitment to adhesions was independent of Fer kinase activity, but was impacted by N-cadherin ligation-provoked Rac activation. We conclude that N-cadherin ligation induces Rac-dependent cortactin recruitment and Fer-dependent cortactin phosphorylation, which in turn promotes enhanced mobilization and interaction of surface expressed N-cadherin in contacting cells.
authors.affiliation: CIHR Group in Matrix Dynamics, University of Toronto, Toronto, Ontario M5S 3E2, Canada.
authors.forename: Tarek Y
authors.initials: TY
authors.lastname: El Sayegh
authors.forename: Pamela D
authors.initials: PD
authors.lastname: Arora
authors.forename: Lingzhi
authors.initials: L
authors.lastname: Fan
authors.forename: Carol A
authors.initials: CA
authors.lastname: Laschinger
authors.forename: Peter A
authors.initials: PA
authors.lastname: Greer
authors.forename: Christopher A
authors.initials: CA
authors.lastname: McCulloch
authors.forename: Andras
authors.initials: A
authors.lastname: Kapus
mesh_terms: D000818:Animals; D015820:Cadherins / Q000502:physiology*; D002448:Cell Adhesion / Q000502:physiology*; D002460:Cell Line; D051356:Cortactin / Q000378:metabolism*; D004789:Enzyme Activation; D020558:GTP Phosphohydrolases / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D051381:Rats
chemical_list: D015820:Cadherins; D051356:Cortactin; C052602:proto-oncogene protein c-fes-fps; D011505:Protein-Tyrosine Kinases; D020558:GTP Phosphohydrolases