CdGAP associates with actopaxin to regulate integrin-dependent changes in cell morphology and motility.
BACKGROUND
Integrin signaling, stimulated by cell adhesion to the extracellular matrix, plays a critical role in coordinating changes in cell morphology and migration. The requisite remodeling of the cytoskeleton is controlled by the Rho family of small GTPases, which are, in turn, regulated via activation by guanine-nucleotide exchange factors (GEFs) and inactivation by GTPase-activating proteins (GAPs). However, the mechanisms contributing to the precise spatial and temporal regulation of these Rho GTPase modulators remain poorly understood.


RESULTS
The Cdc42/Rac GAP CdGAP has previously been implicated as an inhibitor of growth-factor-induced lamellipodia formation. Herein, CdGAP is shown to localize to focal adhesions, potentially through its direct association with the amino terminus of actopaxin, a paxillin and actin binding protein. CdGAP activity is regulated in an adhesion-dependent manner and, through the overexpression of wild-type CdGAP and a GAP-deficient mutant, as well as RNA interference, is shown to be required for normal cell spreading, polarized lamellipodia formation, and cell migration. Introduction of an actopaxin mutant defective for CdGAP binding, or reduction of actopaxin by using RNAi, significantly attenuated these effects.


CONCLUSIONS
We have established that CdGAP is an important regulator of integrin-induced Rho family signaling to the cytoskeleton and that its interaction with the focal-adhesion protein actopaxin is critical for the correct spatial and/or temporal regulation of CdGAP function. A complete understanding of the coordination of signaling events downstream of integrin engagement with the extracellular matrix will provide valuable insight into the regulation of cell migration during processes such as wound repair, development, and tumor cell metastasis.
LaLonde
David P
DP
Department of Cell and Developmental Biology, State University of New York, Upstate Medical University, Syracuse, 13210, USA.
Grubinger
Markus
M
Lamarche-Vane
Nathalie
N
Turner
Christopher E
CE
D000185:Actinin / Q000378:metabolism*; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics / Q000502:physiology*; D016764:Cell Polarity / Q000235:genetics / Q000502:physiology; D048430:Cell Shape / Q000502:physiology; D022001:Focal Adhesions / Q000378:metabolism; D020690:GTPase-Activating Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000032:analysis; D006801:Humans; D016023:Integrins / Q000378:metabolism*; D008164:Luminescent Proteins / Q000032:analysis; D051419:Paxillin / Q000235:genetics / Q000378:metabolism; D000097573:Red Fluorescent Protein
D000185:Actinin; D020690:GTPase-Activating Proteins; D049452:Green Fluorescent Proteins; D016023:Integrins; D008164:Luminescent Proteins; D051419:Paxillin; D000097573:Red Fluorescent Protein