| 1998 |
CdGAP (ARHGAP31) was identified as a novel GTPase-activating protein with in vitro GAP activity toward both Cdc42 and Rac1 but not RhoA. Microinjection of CdGAP into serum-starved fibroblasts inhibited PDGF-induced lamellipodia (Rac-mediated) and bradykinin-induced filopodia (Cdc42-mediated), but had no effect on LPA-induced stress fiber formation. The C-terminus contains potential PKC phosphorylation sites and five SH3 binding motifs. |
Yeast two-hybrid screen, in vitro GAP assay, microinjection into fibroblasts |
The Journal of biological chemistry |
High |
9786927
|
| 2001 |
The endocytic protein intersectin interacts with CdGAP through a subset of its SH3 domains and inhibits CdGAP's GAP activity toward Rac1. In PDGF-stimulated Swiss 3T3 cells, intersectin co-localizes with CdGAP. The central domain of CdGAP (not the proline-rich domain) is required for the CdGAP–intersectin interaction, but the C-terminal proline-rich domain is required for intersectin-mediated inhibition of GAP activity, suggesting a conformational change mechanism. |
Co-immunoprecipitation, in vitro GAP assay with intersectin-SH3 domains, co-localization in PDGF-stimulated cells, deletion mutant analysis |
The Journal of biological chemistry |
High |
11744688
|
| 2005 |
CdGAP is phosphorylated in vivo on serine and threonine residues downstream of the MEK-ERK pathway in response to serum or PDGF. ERK1 and RSK-1 phosphorylate CdGAP in vitro. A DEF domain (docking for ERK, FXFP motif) in the proline-rich region is required for efficient ERK1/2 binding and phosphorylation. Thr776 was identified as an in vivo ERK1/2 target site that serves as an important regulatory site of CdGAP GAP activity. |
In vitro kinase assay, site-directed mutagenesis, phosphorylation mapping, co-immunoprecipitation, cell stimulation with serum/PDGF |
Molecular and cellular biology |
High |
16024771
|
| 2006 |
CdGAP localizes to focal adhesions through a direct interaction with the N-terminus of actopaxin (α-parvin), a paxillin and actin binding protein. CdGAP GAP activity is regulated in an adhesion-dependent manner. Both overexpression and RNAi knockdown of CdGAP impaired normal cell spreading, polarized lamellipodia formation, and cell migration. An actopaxin mutant defective for CdGAP binding attenuated these effects. |
Co-immunoprecipitation/pulldown, RNAi knockdown, overexpression of GAP-deficient mutant, fluorescence localization, cell spreading/migration assays |
Current biology : CB |
High |
16860736
|
| 2006 |
Human CdGAP (KIAA1204) shares 76% sequence identity with mouse CdGAP-long isoform and retains in vitro and in vivo GAP activity toward Cdc42 and Rac1 but not RhoA, confirming substrate specificity is conserved. Human CdGAP is phosphorylated in vivo on serine and threonine residues. Unlike mouse CdGAP, human CdGAP interacts with ERK1/2 through a region not involving a canonical DEF domain. CdGAP expression caused membrane blebbing in COS-7 cells. |
In vitro GAP assay, in vivo phosphorylation analysis, co-immunoprecipitation with ERK1/2, cell morphology assay |
Biology of the cell |
Medium |
16519628
|
| 2006 |
GSK-3α was identified as a CdGAP binding partner. GSK-3α and GSK-3β both interact with CdGAP in mammalian cells and phosphorylate CdGAP in vitro and in vivo at Thr776, the same site previously identified as an ERK1/2 target. GSK-3 activity is necessary for serum-stimulated upregulation of CdGAP protein levels (but not mRNA), revealing a novel mechanism for controlling Cdc42/Rac1 signaling. |
Yeast two-hybrid, co-immunoprecipitation, in vitro kinase assay, in vivo phosphorylation, GSK-3 inhibitor treatment |
The Journal of biological chemistry |
High |
17158447
|
| 2010 |
CdGAP is required for TGF-β-induced cell motility and invasion of Neu/ErbB-2-expressing mammary tumor cells. The proline-rich domain (PRD) but not the GAP domain of CdGAP is essential for TGF-β-mediated motility and invasion. CdGAP depletion increased E-cadherin expression and prevented complete E-cadherin loss during TGF-β-induced EMT. TGF-β also induces CdGAP expression and phosphorylation in NMuMG cells. |
siRNA knockdown, overexpression of deletion mutants (rescue analysis), cell motility/invasion assays, immunoblotting for E-cadherin |
Oncogene |
Medium |
21042277
|
| 2011 |
Gain-of-function truncating mutations in the terminal exon of ARHGAP31 cause Adams-Oliver syndrome. Mutant transcripts are stable and increase ARHGAP31 GAP activity in vitro. Constitutively active ARHGAP31 mutations result in loss of available active Cdc42 and disruption of actin cytoskeletal structures. Mouse Arhgap31 expression is restricted to terminal limb buds and craniofacial processes during early development. |
In vitro GAP activity assay of mutant proteins, genome-wide linkage analysis, exome sequencing, Rho GTPase activity assay (active Cdc42 measurement), mouse expression analysis |
American journal of human genetics |
High |
21565291
|
| 2011 |
The interaction between intersectin's SH3D domain and CdGAP is mediated through a novel basic-rich motif in the central domain of CdGAP containing a conserved xKx(K/R)K sequence, not a canonical proline-rich SH3-binding motif. |
Pulldown assay, deletion and point mutagenesis of CdGAP central domain, direct binding characterization |
FEBS letters |
Medium |
21349274
|
| 2012 |
A polybasic region (PBR) preceding the RhoGAP domain of CdGAP mediates specific binding to PI(3,4,5)P3. In vitro reconstitution with membrane vesicles loaded with prenylated Rac1 showed that the PBR is required for full CdGAP activation in the presence of PI(3,4,5)P3. In fibroblasts, PBR mutants showed reduced ability to induce cell rounding and to negatively regulate cell spreading; the PBR is also required for in vivo Rac1 inactivation. |
In vitro lipid binding assay, membrane vesicle reconstitution with prenylated Rac1 and PI(3,4,5)P3, site-directed mutagenesis, cell spreading assays, Rac1 activity assay |
The Journal of biological chemistry |
High |
22518840
|
| 2012 |
CdGAP negatively regulates directed and random cell migration by controlling adhesion maturation and dynamics (both assembly and disassembly). CdGAP also localizes to adhesions in 3D matrix environments and CdGAP depletion promotes cancer cell migration and invasion through 3D matrices. |
siRNA knockdown, overexpression, live-cell imaging of adhesion dynamics, 3D matrix invasion assays |
Cytoskeleton (Hoboken, N.J.) |
Medium |
22907917
|
| 2014 |
CdGAP is necessary for U2OS osteosarcoma cells to sense extracellular matrix stiffness and undergo durotaxis. CdGAP regulated rigidity-dependent motility by controlling membrane protrusion, adhesion dynamics, and Rac1 activity. CdGAP depletion abolished directed migration toward stiffer substrates. |
siRNA knockdown, PDMS gels with varying stiffness, Rac1 activity assay, adhesion dynamics imaging, durotaxis assay |
PloS one |
Medium |
24632816
|
| 2016 |
CdGAP is required for embryonic vascular development in mice; CdGAP-deficient embryos show impaired vascular development at E15.5, superficial vessel defects, subcutaneous edema, and 44% embryonic/perinatal lethality. CdGAP associates with VEGF receptor-2 and controls VEGF-dependent signaling; CdGAP depletion impairs VEGF-mediated Rac1 activation and reduces phosphorylation of Gab1, Akt, PLCγ, and SHP2. |
CdGAP knockout mice, co-immunoprecipitation (CdGAP–VEGFR2), Rac1 activity assay, phosphorylation analysis by immunoblot, aortic ring sprouting assay, endothelial cell migration assay |
Scientific reports |
High |
27270835
|
| 2017 |
CdGAP uses its proline-rich domain to form a functional complex with Zeb2 to mediate transcriptional repression of E-cadherin (CDH1) in ErbB2-transformed breast cancer cells, acting as a nuclear transcriptional co-repressor in a GAP-independent manner. CdGAP knockdown decreased Snail1 and Zeb2 levels, increased E-cadherin, and restored cell-cell junctions. In vivo, CdGAP loss impaired tumor growth and lung metastasis. |
Co-immunoprecipitation (CdGAP–Zeb2), gene expression profiling, reporter assays for E-cadherin promoter, deletion mutant analysis, in vivo mouse tumor/metastasis model |
Oncogene |
High |
28135249
|
| 2017 |
Ajuba scaffold protein interacts with CdGAP at cell-cell contacts and inhibits CdGAP GAP activity. CdGAP recruitment to junctions does not require Ajuba; rather Ajuba controls CdGAP residence at cell-cell contacts. Ajuba binds Rac1 and CdGAP via distinct domains and can bring them into proximity to regulate Rac1 activity. Gain-of-function AOS CdGAP mutants strongly destabilize cell-cell contacts. |
Co-immunoprecipitation, GAP activity assay, fluorescence localization, overexpression of Ajuba/CdGAP mutants, junction integrity assays |
Scientific reports |
Medium |
28835688
|
| 2018 |
RSK phosphorylates CdGAP at Ser1093 and Ser1163 in response to phorbol ester, creating docking sites for 14-3-3 adaptor proteins. Binding of 14-3-3β to CdGAP inhibits CdGAP GAP activity, sequesters CdGAP in the cytoplasm, inhibits nucleocytoplasmic shuttling, abolishes CdGAP-induced cell rounding, inhibits CdGAP-mediated E-cadherin promoter repression, and inhibits CdGAP-induced cell migration. AOS-related CdGAP mutant proteins lacking these phospho-residues are not regulated by 14-3-3β. |
In vitro kinase assay (RSK), phospho-site mutagenesis, co-immunoprecipitation (CdGAP–14-3-3β), GAP activity assay, subcellular fractionation, cell migration assay, E-cadherin promoter reporter assay |
Oncotarget |
High |
29545927
|
| 2022 |
CdGAP interacts with β-PIX (ARHGEF7) through its basic region in podocytes. Upon EGF stimulation, both proteins translocate to the plasma membrane together. CdGAP depletion results in altered podocyte motility, increased basal Rac1 and Cdc42 activities, impaired β-PIX membrane translocation and tyrosine phosphorylation, and reduced activities of Src kinase, FAK, and paxillin. Podocyte-specific CdGAP knockout mice develop mild proteinuria exacerbated by Adriamycin. |
Proximity-based ligation assay, co-immunoprecipitation, siRNA knockdown, live imaging, GTPase activity assay, podocyte-specific knockout mice |
Scientific reports |
Medium |
36333327
|
| 2023 |
DLG1 transcriptionally regulates ARHGAP31 expression, and this DLG1-ARHGAP31-CDC42 axis is essential for intestinal stem cell (ISC) survival in the context of increased Wnt signaling. ARHGAP31 acts downstream of DLG1 to deactivate CDC42, an effector of the non-canonical Wnt pathway, thereby modulating ISC response to fluctuating niche Wnt levels. |
RNA sequencing, genetic mouse models (conditional knockouts), epistasis analysis, intestinal organoids |
Cell stem cell |
Medium |
36640764
|
| 2023 |
CdGAP interacts with the adaptor protein talin to modulate focal adhesion dynamics and integrin activation. CdGAP is positively regulated by TGF-β signaling during EMT and promotes tumor formation and metastasis in HER2+ breast cancer. CdGAP depletion mediates crosstalk with a DLC1-RhoA pathway in HER2+ primary tumors and is associated with a TGF-β-induced EMT transcriptional signature. |
Co-immunoprecipitation (CdGAP–talin), focal adhesion dynamics imaging, in vivo HER2+ mouse breast cancer model, gene expression profiling, intravasation/extravasation assays |
Cell reports |
Medium |
37552602
|