title: A non-canonical role for Rgnef in promoting integrin-stimulated focal adhesion kinase activation.
abstract: Rgnef (also known as p190RhoGEF or ARHGEF28) is a Rho guanine-nucleotide-exchange factor (GEF) that binds focal adhesion kinase (FAK). FAK is recruited to adhesions and activated by integrin receptors binding to matrix proteins, such as fibronectin (FN). Canonical models place Rgnef downstream of integrin-FAK signaling in regulating Rho GTPase activity and cell movement. Herein, we establish a new, upstream role for Rgnef in enhancing FAK localization to early peripheral adhesions and promoting FAK activation upon FN binding. Rgnef-null mouse embryo fibroblasts (MEFs) exhibit defects in adhesion formation, levels of FAK phosphotyrosine (pY)-397 and FAK localization to peripheral adhesions upon re-plating on FN. Rgnef re-expression rescues these defects, but requires Rgnef-FAK binding. A mutation in the Rgnef pleckstrin homology (PH) domain inhibits adhesion formation, FAK localization, and FAK-Y397 and paxillin-Y118 phosphorylation without disrupting the Rgnef-FAK interaction. A GEF-inactive Rgnef mutant rescues FAK-Y397 phosphorylation and early adhesion localization, but not paxillin-Y118 phosphorylation. This suggests that, downstream of FN binding, paxillin-pY118 requires Rgnef GEF activity through a mechanism distinct from adhesion formation and FAK activation. These results support a scaffolding role for Rgnef in FAK localization and activation at early adhesions in a PH-domain-dependent but GEF-activity-independent manner.
authors.affiliation: Moores UCSD Cancer Center, La Jolla, CA 92093, USA.
authors.forename: Nichol L G
authors.initials: NL
authors.lastname: Miller
authors.forename: Christine
authors.initials: C
authors.lastname: Lawson
authors.forename: Elizabeth G
authors.initials: EG
authors.lastname: Kleinschmidt
authors.forename: Isabelle
authors.initials: I
authors.lastname: Tancioni
authors.forename: Sean
authors.initials: S
authors.lastname: Uryu
authors.forename: David D
authors.initials: DD
authors.lastname: Schlaepfer
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002448:Cell Adhesion; D002478:Cells, Cultured; D004789:Enzyme Activation; D005347:Fibroblasts / Q000166:cytology / Q000201:enzymology / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019012:Integrin beta1 / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D008969:Molecular Sequence Data; D051419:Paxillin / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment; D015398:Signal Transduction; D020666:ras-GRF1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D005353:Fibronectins; D019012:Integrin beta1; D051419:Paxillin; C513037:Rasgrf1 protein, mouse; D020666:ras-GRF1; D051416:Focal Adhesion Protein-Tyrosine Kinases
keywords: Adhesion; Arhgef28; FAK; GEF; PH domain; Rgnef; p190RhoGEF