title: The small GTPase Rap1 promotes cell movement rather than stabilizes adhesion in epithelial cells responding to insulin-like growth factor I.
abstract: The Ras-related GTPase Rap1 promotes cell adhesion and migration. Although the significance of Rap1 contribution to cell migration is increasingly being recognized, little is known about the biochemical mechanisms driving this process. In the present study, we discovered a previously unidentified regulatory role of insulin-like growth factor type I (IGF-I) receptor (IGF-IR) in CRK Src homology 3 (SH3)-binding guanine-nucleotide-releasing protein (C3G)-Rap1-fascin-actin axis promoting cell movement. We demonstrate that a burst of Rap1 activity, rather than presumed hyperactivation, is imperative for the onset of cell movement. We show that while autophosphorylated IGF-IR signals to C3G to activate Rap1, subsequent IGF-IR internalization promotes gradual inactivation of Rap1 by putative Rap1 GTPase-activating protein (GAP). Additionally, IGF-IR signalling recruits active Rap1 at sites of cell motile protrusions. C3G depletion prevents IGF-I-induced fascin accumulation at actin microspikes and blocks protrusions. In the absence of IGF-IR activity, the wild-type (WT) Rap1 and the constitutively active V12Rap1 mutant remain in cell-cell contacts. Forced inactivation of Rap1 signalling by overexpressing dominant negative N17Rap1, Rap1GAP or by silencing C3G has a detrimental effect on filamentous (F)-actin and cell adhesion irrespective of IGF-IR signalling. We conclude that the basal levels of Rap1 activity holds up cell adhesion, whereas sequential regulation of C3G and GAP by IGF-IR reverses the labile Rap1 function from supporting adhesion to promoting migration.
authors.forename: Marina A
authors.initials: MA
authors.lastname: Guvakova
authors.affiliation: *Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, U.S.A.
authors.forename: William S Y
authors.initials: WS
authors.lastname: Lee
authors.affiliation: †Department of Surgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, U.S.A.
authors.forename: Dana K
authors.initials: DK
authors.lastname: Furstenau
authors.affiliation: †Department of Surgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, U.S.A.
authors.forename: Indira
authors.initials: I
authors.lastname: Prabakaran
authors.affiliation: †Department of Surgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, U.S.A.
authors.forename: David C
authors.initials: DC
authors.lastname: Li
authors.affiliation: †Department of Surgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, U.S.A.
authors.forename: Rupert
authors.initials: R
authors.lastname: Hung
authors.affiliation: *Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, U.S.A.
authors.forename: Natasha
authors.initials: N
authors.lastname: Kushnir
mesh_terms: D002448:Cell Adhesion; D002465:Cell Movement; D004847:Epithelial Cells / Q000166:cytology / Q000201:enzymology* / Q000378:metabolism; D020735:Guanine Nucleotide-Releasing Factor 2 / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000235:genetics / Q000378:metabolism*; D061986:MCF-7 Cells; D017451:Receptors, Somatomedin / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D020827:rap1 GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D020735:Guanine Nucleotide-Releasing Factor 2; D017451:Receptors, Somatomedin; D007334:Insulin-Like Growth Factor I; D020827:rap1 GTP-Binding Proteins