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

TheRas-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. Here, we discovered a previously unidentified regulatory role of insulin-like growth factor type I receptor (IGF-IR) in CRK SH3-binding guanine nucleotide exchange factor (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 Rap1GTPase-activating protein (GAP). Additionally, IGF-IR signaling 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’s activity, the wild type Rap1 and the constitutively active V12Rap1 mutant remain in cell-cell contacts. Forced inactivation of Rap1 signaling by overexpressing dominant negative N17Rap1, Rap1GAP, or by silencing C3G has a detrimental effect on F-actin and cell adhesion irrespectively of IGF-IR signaling. We conclude that the basal levels of Rap1 activity hold up cell adhesion, whereas sequential regulation of C3G and GAP by IGF-IR reverses the labile Rap1 function from suppo...

Source: BJ Cell - July 16, 2014 Category: Biochemistry Authors: M A Guvakova, W S.Y. Lee, D K Furstenau, I Prabakaran, D C. Li, R Hung, N Kushnir Tags: BJ Cell Source Type: research