The small GTPase Rap1 promotes cell movement rather than stabilizes adhesion in epithelial cells responding to insulin-like growth factor I

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 re...

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Bibliographic Details
Published in:	Biochemical journal Vol. 463; no. 2; p. 257
Main Authors:	Guvakova, Marina A, Lee, William S Y, Furstenau, Dana K, Prabakaran, Indira, Li, David C, Hung, Rupert, Kushnir, Natasha
Format:	Journal Article
Language:	English
Published:	England 15-10-2014
Subjects:	Cell Adhesion Cell Movement Epithelial Cells - cytology Epithelial Cells - enzymology Epithelial Cells - metabolism Guanine Nucleotide-Releasing Factor 2 - genetics Guanine Nucleotide-Releasing Factor 2 - metabolism Humans Insulin-Like Growth Factor I - genetics Insulin-Like Growth Factor I - metabolism MCF-7 Cells rap1 GTP-Binding Proteins - genetics rap1 GTP-Binding Proteins - metabolism Receptors, Somatomedin - genetics Receptors, Somatomedin - metabolism Signal Transduction
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Summary:	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.
ISSN:	1470-8728
DOI:	10.1042/BJ20131638