title: Focal adhesion kinase is required for the spatial organization of the leading edge in migrating cells.
abstract: The process of cell migration is initiated by protrusion at the leading edge of the cell, the formation of peripheral adhesions, the exertion of force on these adhesions, and finally the release of the adhesions at the rear of the cell. Focal adhesion kinase (FAK) is intimately involved in the regulation of this process, although the precise mechanism(s) whereby FAK regulates cell migration is unclear. We have used two approaches to reduce FAK expression in fibroblasts. Treatment of cells with FAK-specific siRNAs substantially reduced FAK expression and inhibited the spreading of fibroblasts in serum-free conditions, but did not affect the rate of spreading in the presence of serum. In contrast with the wild-type cells, the FAK siRNA-treated cells exhibited multiple extensions during cell spreading. The extensions appeared to be inappropriately formed lamellipodia as evidenced by the localization of cortactin to lamellipodial structures and the inhibition of such structures by expression of dominant-negative Rac. The wild-type phenotype was restored by reexpressing wild-type FAK in the knockdown cells, but not by expression of FAK containing a point mutation at the autophosphorylation site (FAK Y397F). In wound-healing assays, FAK knockdown cells failed to form broad lamellipodia, instead forming multiple leading edges. Similar results were obtained using primary mouse embryo fibroblasts from FAK-flox mice in which Cre-mediated excision was used to ablate the expression of FAK. These data are consistent with a role for FAK in regulating the formation of a leading edge during cell migration by coordinating integrin signaling to direct the correct spatial activation of membrane protrusion.
authors.affiliation: Department of Microbiology, University of Virginia Health System, Charlottesville, 22908, USA.
authors.forename: Robert W
authors.initials: RW
authors.lastname: Tilghman
authors.forename: Jill K
authors.initials: JK
authors.lastname: Slack-Davis
authors.forename: Natalia
authors.initials: N
authors.lastname: Sergina
authors.forename: Karen H
authors.initials: KH
authors.lastname: Martin
authors.forename: Marcin
authors.initials: M
authors.lastname: Iwanicki
authors.forename: E Daniel
authors.initials: ED
authors.lastname: Hershey
authors.forename: Hilary E
authors.initials: HE
authors.lastname: Beggs
authors.forename: Louis F
authors.initials: LF
authors.lastname: Reichardt
authors.forename: J Thomas
authors.initials: JT
authors.lastname: Parsons
mesh_terms: D000818:Animals; D002460:Cell Line; D002465:Cell Movement; D016764:Cell Polarity / Q000235:genetics; D048430:Cell Shape; D005347:Fibroblasts / Q000166:cytology / Q000201:enzymology / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; D022001:Focal Adhesions; D017353:Gene Deletion; D005786:Gene Expression Regulation; D006056:Golgi Apparatus / Q000378:metabolism; D051379:Mice; D011505:Protein-Tyrosine Kinases / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D011554:Pseudopodia; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D051381:Rats; D013997:Time Factors
chemical_list: D005353:Fibronectins; D034741:RNA, Small Interfering; D011505:Protein-Tyrosine Kinases; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; C496051:Ptk2 protein, mouse; C496052:Ptk2 protein, rat