title: Overexpression of FAK promotes Ras activity through the formation of a FAK/p120RasGAP complex in malignant astrocytoma cells.
abstract: Focal adhesion kinase (FAK) signaling may be mediated through the modulation of Ras activity. We have shown previously that grade III malignant astrocytoma biopsy samples exhibit elevated levels of FAK, and that overexpression of FAK in U-251MG malignant astrocytoma cells promotes the phosphorylation of Shc, a potential upstream mediator of Ras activity. Here, we report that overexpression of FAK promotes Ras activity in U-251MG malignant astrocytoma cells cultured in aggregate suspension or as monolayers adherent to vitronectin. The overexpression of FAK also promoted the association of FAK with p120RasGAP, which is a negative regulator of Ras activity, in the U-251MG cells cultured in aggregate suspension, with this association being abrogated upon plating of the cells onto vitronectin. An association of FAK with p120RasGAP also was observed in malignant astrocytoma biopsy samples, but not in normal brain samples. As overexpression of FAK in U-251MG cells in aggregate suspension culture reduced the amount of p120RasGAP complexed with active Ras, we hypothesize that the association of FAK with p120 RasGAP may facilitate Ras activity. The overexpression of a mutated FAK in which the Y397 had been mutated to F did not result in the formation of the FAK/p120RasGAP complex and did not promote Ras activity, indicating that the Y397 residue of FAK plays a role in the formation of this complex and in the activation of Ras. Moreover, the overexpression of mutated FAK (397F) was found to inhibit anchorage-independent growth. These data provide the basis for a previously undescribed mechanism in which the elevated expression of FAK can promote Ras activity through its competitive recruitment of p120RasGAP, thereby diminishing the association of p120RasGAP with active Ras.
authors.affiliation: Department of Pathology, Division of Neuropathology, The University of Alabama at Birmingham, Birmingham, AL 35294, USA.
authors.forename: Timothy P
authors.initials: TP
authors.lastname: Hecker
authors.forename: Qiang
authors.initials: Q
authors.lastname: Ding
authors.forename: Tanya A
authors.initials: TA
authors.lastname: Rege
authors.forename: Steven K
authors.initials: SK
authors.lastname: Hanks
authors.forename: Candece L
authors.initials: CL
authors.lastname: Gladson
mesh_terms: D000818:Animals; D001254:Astrocytoma / Q000378:metabolism*; D002455:Cell Division / Q000502:physiology; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; D006160:Guanosine Triphosphate / Q000378:metabolism; D006801:Humans; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D051381:Rats; D020729:p120 GTPase Activating Protein / Q000378:metabolism*; D018631:ras Proteins / Q000378:metabolism*
chemical_list: D020729:p120 GTPase Activating Protein; D006160:Guanosine Triphosphate; D011505:Protein-Tyrosine Kinases; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; C496050:PTK2 protein, human; C496052:Ptk2 protein, rat; D018631:ras Proteins