Affinage

ARHGDIA

Rho GDP-dissociation inhibitor 1 · UniProt P52565

Length
204 aa
Mass
23.2 kDa
Annotated
2026-06-09
100 papers in source corpus 41 papers cited in narrative 40 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 7/7 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

ARHGDIA (RhoGDIα) is a cytosolic regulator that maintains prenylated Rho-family GTPases (RhoA, Rac1, Cdc42) in an inactive, soluble pool and shapes the spatiotemporal availability of active GTPases at membranes (PMID:11149925, PMID:31647414). It engages its GTPase clients through two cooperating interfaces resolved by crystallography and NMR: a flexible N-terminal regulatory arm that contacts the switch I/II regions to block GDP dissociation and GAP-stimulated GTP hydrolysis, and a structured immunoglobulin-like C-terminal domain whose hydrophobic pocket sequesters the geranylgeranyl group to mediate membrane extraction (PMID:10676816, PMID:10489445, PMID:9195882, PMID:10673424, PMID:11114252). RhoGDI binds GDP-bound RhoA with extraordinary affinity (Kd ~5 pM) and clamps even GTP-loaded GTPase into a GDP-like conformation without hydrolysis, making membrane extraction a thermodynamically favored passive process (PMID:22628549); the Rho insert region and switch II arginines of the GTPase are required for this functional inhibition independent of binding (PMID:12956948, PMID:9334181). Client release is gated by multisite phosphorylation of RhoGDI — PAK1 at Ser101/Ser174 selectively frees Rac1 (PMID:15225553), Src at Tyr156 releases RhoA/Rac1/Cdc42 (PMID:16943322), and PKCα at Ser96 releases RhoA and Rac1 (PMID:23776598) — and by displacement factors including the p75NTR receptor, whose PKC-phosphorylation-dependent interaction with the RhoGDI N-terminus liberates RhoA to restrain neurite outgrowth (PMID:12692556, PMID:38253689). Membrane phosphoinositides acting together with GEF-mediated nucleotide exchange also dissociate Rac1·RhoGDI complexes (PMID:18505730), and PKA-mediated phosphorylation of RhoA conversely strengthens its capture by RhoGDI to pace protrusion-retraction cycles in migrating cells (PMID:21572420). RhoGDI binding affinity and stability are further tuned by SUMOylation at Lys-138 and by competing ubiquitination (PMID:22393046, PMID:31362179), and RhoGDI cooperates with the KLHL23-Cul3 E3 ligase to inactivate Cdc42 through complementary GDP- and GTP-state targeting (PMID:40846997). Loss-of-function ARHGDIA mutations cause steroid-resistant nephrotic syndrome through uncontrolled activation of RAC1 and CDC42 in podocytes (PMID:23867502, PMID:23434736).

Mechanistic history

Synthesis pass · year-by-year structured walk · 11 steps
  1. 1993 High

    Established that RhoGDI is not merely a GDP-dissociation inhibitor but can bind both GDP- and GTP-loaded GTPases and shield active Rac from GAP inactivation, defining its biochemical scope.

    Evidence In vitro reconstitution with recombinant Rho/Rac, GAPs, and prenylation/processing mutants

    PMID:8491184

    Open questions at the time
    • Did not resolve the structural basis of two-interface binding
    • Physiological role of GTP-state binding unaddressed
  2. 1997 High

    Defined the bipartite architecture of RhoGDI — a flexible N-terminal arm essential for binding and a C-terminal Ig-like hydrophobic pocket for the isoprenyl group — and showed both are needed for full activity.

    Evidence Crystallography, NMR, and selective proteolysis of RhoGDI; chimeric Cdc42 Rho-insert mapping

    PMID:9195882 PMID:9334181

    Open questions at the time
    • Did not capture the GTPase-bound complex
    • Sequence elements distinguishing binding from functional inhibition only partly resolved
  3. 2000 High

    Crystal structures of GTPase·RhoGDI complexes revealed the two-site mechanism: N-terminal arm occludes switch regions and the GEF epitope while the geranylgeranyl group inserts into the Ig-like pocket, explaining nucleotide-exchange inhibition and membrane release.

    Evidence X-ray structures of Cdc42/RhoGDI, RhoA/RhoGDI, and Rac1/RhoGDI; NMR mapping of Rac binding site

    PMID:10489445 PMID:10673424 PMID:10676816 PMID:11513578

    Open questions at the time
    • Static structures did not address kinetics of release
    • Did not explain how active GTPases escape sequestration
  4. 2001 High

    Demonstrated in cells that RhoGDI binding sequesters specific Rho GTPases in the cytosol and controls their membrane targeting, with binding lost on constitutively active/dominant-negative mutants and on certain GTPases (RhoB, TC10).

    Evidence GFP live-cell imaging, Co-IP, palmitoylation insertion, and mutagenesis across multiple GTPases

    PMID:11149925 PMID:11583574

    Open questions at the time
    • Selectivity determinants for excluded GTPases not fully defined
    • Mechanism of cytosol-to-membrane cycling left open
  5. 2004 High

    Identified PAK1 phosphorylation of RhoGDI at Ser101/Ser174 as a Rac1-selective release switch, providing the first kinase-controlled mechanism for client-specific dissociation.

    Evidence In vitro and in vivo kinase assays, Co-IP, and site-specific mutagenesis

    PMID:15225553

    Open questions at the time
    • How selectivity for Rac1 over RhoA arises structurally not resolved
    • Upstream signals controlling PAK1 not addressed here
  6. 2006 High

    Showed Src phosphorylation at Tyr156 broadly dissociates all three GTPase clients and drives RhoGDI to the membrane/cortical actin, linking tyrosine kinase signaling to global Rho activation.

    Evidence In vitro kinase assay, Co-IP, fractionation, and phosphomimetic imaging

    PMID:16943322

    Open questions at the time
    • Membrane recruitment partner for phospho-RhoGDI unidentified
    • Integration with PAK1/PKC phosphorylation events unresolved
  7. 2008 High

    Reconstituted release in vitro, establishing that anionic phosphoinositide membranes plus a Rac GEF and GTP cooperatively dissociate Rac1·RhoGDI, defining the membrane-level requirements for activation.

    Evidence Fully reconstituted system with prenylated Rac1·RhoGDI, defined liposomes, GEFs, and NADPH oxidase readout

    PMID:18505730

    Open questions at the time
    • In-cell sufficiency of this pathway not tested here
    • Quantitative contribution relative to phosphorylation routes unknown
  8. 2012 High

    Quantified the thermodynamics of extraction, showing RhoGDI binds GDP-RhoA with pM affinity, forces GTP-RhoA into a GDP-like conformation, and extracts from membranes passively through tightening intermediates.

    Evidence Quantitative fluorescence binding kinetics and X-ray structure of RhoA·GMPPNP·RhoGDI

    PMID:22628549

    Open questions at the time
    • How displacement factors overcome pM affinity in vivo not addressed
    • Kinetic intermediates not structurally captured
  9. 2019 High

    Established RhoGDI as an active spatial patterning factor that extracts not only inactive but also active GTPases from membranes to sculpt zones of activity, e.g. around cell wounds.

    Evidence In vivo Xenopus fluorescence imaging and in vitro reconstitution on lipid bilayers

    PMID:31647414

    Open questions at the time
    • Regulatory inputs selecting active-state extraction unclear
    • Generalizability beyond wound contexts untested
  10. 2013 High

    Connected RhoGDI directly to human disease, showing ARHGDIA loss-of-function mutations cause steroid-resistant nephrotic syndrome via hyperactivation of RAC1 and CDC42 in podocytes.

    Evidence Whole-exome sequencing, Co-IP, GTPase activity assays, zebrafish models, and RAC1 inhibitor rescue

    PMID:23434736 PMID:23867502

    Open questions at the time
    • Why RHOA is spared in patient mutants not mechanistically explained
    • Podocyte-specific downstream effectors not fully defined
  11. 2025 High

    Revealed coordinated control of Cdc42 by RhoGDI and the KLHL23-Cul3 ligase, which compete for switch II in a nucleotide-state-selective manner (RhoGDI for GDP, KLHL23 for GTP) to inactivate Cdc42 spatiotemporally.

    Evidence FRET, ubiquitylation assays, KLHL23 depletion, disease-variant analysis, and metastasis models

    PMID:40846997

    Open questions at the time
    • How RhoGDI and KLHL23 hand off Cdc42 dynamically not resolved
    • Whether analogous ligase coordination applies to RhoA/Rac1 unknown

Open questions

Synthesis pass · forward-looking unresolved questions
  • How the many regulatory inputs — multisite phosphorylation, SUMO/ubiquitin competition, phosphoinositide/GEF action, displacement receptors, and competing E3 ligases — are integrated to produce client- and location-specific GTPase activation in a given cell remains unresolved.
  • No unified quantitative model integrating phosphorylation, lipid, and ligase inputs
  • Cell-type-specific hierarchy of release mechanisms unknown

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0098772 molecular function regulator activity 4 GO:0140313 molecular sequestering activity 4 GO:0008289 lipid binding 3 GO:0140096 catalytic activity, acting on a protein 2
Localization
GO:0005829 cytosol 4 GO:0005886 plasma membrane 3
Pathway
R-HSA-162582 Signal Transduction 4 R-HSA-1643685 Disease 3
Partners
CDC42KLHL23NGFRPAK1RAC1RHOASRCXIAP
Complex memberships
DGKzeta-PAK1-RhoGDI complexRhoGDI-Cdc42 heterodimerRhoGDI-Rac1 heterodimerRhoGDI-RhoA heterodimer

Evidence

Reading pass · 40 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2000 Crystal structure of Cdc42/RhoGDI complex at 2.6 Å reveals two interaction sites: (1) the N-terminal regulatory arm of RhoGDI binds switch I and II domains of Cdc42, inhibiting GDP dissociation and GTP hydrolysis; (2) the geranylgeranyl moiety of Cdc42 inserts into a hydrophobic pocket in the immunoglobulin-like domain of RhoGDI, mediating membrane release. X-ray crystallography Cell High 10676816
2001 Crystal structure of Rac1/RhoGDI complex at 2.7 Å reveals geranylgeranyl moiety of Rac1 inserts into RhoGDI, causing structural changes in the RhoGDI core; Rho insert region residues Tyr64, Arg66, His103, His104, Leu67, Leu70 mediate protein-protein contacts; inhibition of GDP-GTP exchange on Rac1 results partly from interaction of Thr35(Rac) with Asp45(GDI); effector loops of Rac1 remain accessible in the complex, enabling NADPH oxidase activation. X-ray crystallography Biochemistry High 11513578
1999 Crystal structure of the RhoA-RhoGDI complex shows the N-terminus of RhoGDI binds switch I and switch II regions of RhoA, occluding the epitope for Dbl-like nucleotide exchange factors; the hydrophobic pocket entrance is oriented to accommodate the geranylgeranyl group of RhoA. X-ray crystallography (MAD + MIR phasing) Acta crystallographica Section D High 10489445
2012 Quantitative binding analysis shows RhoGDI binds prenylated RhoA•GDP with very high affinity (Kd ~5 pM) and prenylated RhoA•GTP with ~600-fold lower affinity (Kd ~3 nM); 2.8 Å structure of RhoA•GMPPNP•RhoGDI complex reveals that RhoGDI forces activated RhoA into a GDP-bound conformation without nucleotide hydrolysis; membrane extraction by RhoGDI is a thermodynamically favored passive process proceeding through progressively tighter intermediates. Quantitative fluorescence binding assay, X-ray crystallography The Journal of biological chemistry High 22628549
1997 NMR spectroscopy and X-ray crystallography reveal RhoGDI has an N-terminal flexible arm (first ~50-60 residues) essential for binding Rac, and a structured C-terminal immunoglobulin-like domain with an unusual hydrophobic pocket between beta sheets that binds the isoprenyl group of Rac; both domains are required for full activity. X-ray crystallography + NMR spectroscopy + selective proteolysis Structure High 9195882
2004 Pak1 binds and phosphorylates RhoGDI at Ser101 and Ser174 both in vitro and in vivo; this phosphorylation selectively dissociates Rac1-RhoGDI complexes but not RhoA-RhoGDI complexes; Pak1 autoinhibitory domain blocks Cdc42-induced Rac1 activation and PDGF/EGF-stimulated Rac1-RhoGDI dissociation in a manner dependent on Ser101/Ser174 phosphorylation. In vitro kinase assay, in vivo phosphorylation, coimmunoprecipitation, complexation assays Molecular cell High 15225553
2006 Src kinase binds and phosphorylates RhoGDI at Tyr156 in vitro and in vivo; Src-mediated phosphorylation dramatically decreases RhoGDI's ability to complex with RhoA, Rac1, or Cdc42; phosphomimetic Y156E mutant constitutively associates with plasma membrane/cortical actin; expression of RhoGDI(Y156E) enhances cell spreading and membrane ruffling. In vitro kinase assay, coimmunoprecipitation, cell fractionation, fluorescence microscopy Molecular biology of the cell High 16943322
2001 RhoGDI binding to Rho GTPases (RhoA, Rac1, Rac2, Cdc42hs) sequesters them in the cytosol and regulates their membrane targeting; constitutively active or dominant-negative mutations in RhoA, Cdc42hs, or Rac1 abrogate RhoGDI binding and redirect localization to plasma membranes and internal membranes; a palmitoylation site inserted into RhoA blocks RhoGDI binding; RhoB and TC10 are not regulated by RhoGDI. GFP live-cell imaging, coimmunoprecipitation, palmitoylation inhibition, site-directed mutagenesis The Journal of cell biology High 11149925
2003 p75NTR directly interacts with RhoGDI; this interaction is strengthened by MAG or Nogo; p75NTR acts as a displacement factor that releases prenylated RhoA from RhoGDI, thereby activating RhoA and inhibiting axonal elongation; a peptide corresponding to the fifth alpha-helix of p75NTR inhibits p75NTR-RhoGDI interaction. Coimmunoprecipitation, in vitro binding, cell biology assays Nature neuroscience High 12692556
1994 In neutrophils, p21rac2 exists almost entirely complexed with RhoGDI as a 45-50 kDa heterodimer; upon activation (PMA, fMLP, or SDS), a fraction of p21rac2 dissociates from RhoGDI and translocates to the plasma membrane alongside p47phox and p67phox; the rac/GDI complex is active in the cell-free NADPH oxidase assay; GTP enhances and GDP inhibits superoxide production. Cell fractionation, western blotting, cell-free NADPH oxidase assay The Biochemical journal High 8141770
1993 RhoGDI can form stable complexes with Rho and Rac proteins in both GTP- and GDP-bound states; geranylgeranylation and AAX proteolysis are required for efficient RhoGDI interaction, but methylesterification is not; the Rac-GTP–RhoGDI complex is resistant to GAP-stimulated (RhoGAP, BCR) GTP hydrolysis, suggesting RhoGDI can protect active Rac from GAP inactivation. In vitro binding assays, GTPase activity assays with recombinant proteins The EMBO journal High 8491184
2002 Integrin-mediated adhesion promotes translocation of GTP-Rac to membranes via the polybasic C-terminal sequence; cytoplasmic GTP-Rac bound to RhoGDI cannot interact with effectors; release of RhoGDI upon membrane translocation allows Rac to bind effectors; FRET assays show Rac-effector interactions are spatially restricted near cell edges after integrin stimulation. FRET-based live-cell assay, cell fractionation, fluorescence microscopy Nature cell biology High 11862216
2011 PKA phosphorylates RhoA, which increases RhoA interaction with RhoGDI; this establishes a negative feedback loop at the leading edge that controls cycling of RhoA activity and governs the protrusion-retraction pacemaker in migrating cells; live biosensor imaging shows PKA activity closely synchronizes with RhoA activity and membrane protrusion. Live-cell imaging with biosensors, FRET, correlative image analysis Nature cell biology High 21572420
2003 X-ray crystallographic analysis of Cdc42-RhoGDI complex identified Arg66 and Arg68 in the switch II domain of Cdc42 as essential for RhoGDI binding; R66A mutation in constitutively active Cdc42(F28L) abrogated RhoGDI binding without affecting other regulators/effectors; RhoGDI-binding-defective Cdc42(F28L,R66A) was transformation-defective; RhoGDI siRNA inhibited Cdc42(F28L)-mediated transformation, demonstrating RhoGDI is required for Cdc42-mediated cellular transformation. X-ray crystallography, mutagenesis, soft-agar colony assay, RNAi Current biology High 12956948
2009 RhoGDI is identified as a Cdc42 cycling factor in pancreatic beta cells; RhoGDI knockdown selectively amplifies second-phase insulin secretion; glucose stimulation induces tyrosine phosphorylation of RhoGDI causing RhoGDI-Cdc42 complex dissociation at 3 min; subsequent serine phosphorylation causes RhoGDI-Rac1 dissociation; Y156F mutation blocks RhoGDI-Cdc42 dissociation, while Y156F/S101A/S174A triple mutant inhibits the second/granule mobilization phase of insulin secretion. Tandem affinity purification-MS, RNAi, coimmunoprecipitation, mutagenesis, insulin secretion assay The Journal of biological chemistry High 20028975
2013 ARHGDIA mutations (R120X and G173V) identified in steroid-resistant nephrotic syndrome abrogate interaction with RHO GTPases and increase active GTP-bound RAC1 and CDC42 (but not RHOA) in podocytes; a separate in-frame deletion (p.Asp185del) likewise abolishes binding to RhoA, Rac1, and Cdc42; knockdown of ARHGDIA in podocytes hyperactivates all three Rho GTPases and impairs podocyte motility; RAC1 inhibitors partially reverse arhgdia-deficient zebrafish phenotype. Whole-exome sequencing, coimmunoprecipitation, GTPase activity assays, zebrafish knockout, RAC1 inhibitor rescue The Journal of clinical investigation High 23434736 23867502
2007 RhoGDI-1 knockout mice show 2-fold increased basal pulmonary microvascular permeability with opening of interendothelial junctions; RhoA activity (but not Rac1 or Cdc42) is specifically elevated in RhoGDI-1(-/-) lungs and in siRNA-depleted endothelial cells; RhoGDI-1 modulates endothelial barrier function by repressing RhoA activity in vivo. RhoGDI-1(-/-) mice, GTPase pull-down activity assay, siRNA, permeability measurement, electron microscopy Circulation research High 17525371
2019 RhoGDI can extract both inactive (GDP-bound) and active (GTP-bound) RhoGTPases from membranes; extraction of active RhoGTPase by RhoGDI contributes to the spatial regulation of RhoGTPases around cell wounds; direct visualization in vivo and reconstitution on lipid bilayers in vitro establish RhoGDI as an active contributor to spatiotemporal patterning of RhoGTPases. In vivo fluorescence imaging (Xenopus), reconstitution on lipid bilayers in vitro eLife High 31647414
2001 NMR spectroscopy and mutagenesis map the Rac1 binding site on RhoGDI-1: residues 46-57 of the flexible N-terminal domain (forming a transient amphipathic helix) contribute substantially to binding energy; the folded domain engages Rac1 through beta4-beta5 and beta6-beta7 loops; isoprenyl group of Rac1 binds in a distinct pocket on the same face; three distinct interaction sites on RhoGDI collectively bind isoprenylated Rac1. NMR spectroscopy, site-directed mutagenesis Structure High 10673424
2001 NMR characterization of RhoGDI N-terminal domain identifies two regions with helical tendency (residues 36-58 and 9-20); truncation studies show first 30 residues are not required for GDP dissociation inhibition but are important for GTP hydrolysis inhibition; removal of first 41 residues completely abolishes inhibition of GDP dissociation; these structural findings explain why RhoGDI and D4GDI differ in their ability to regulate GTP-bound forms of Rho GTPases. NMR spectroscopy (15N relaxation), truncation mutants, in vitro functional assays Journal of molecular biology High 11114252
1998 RhoGDI associates with a Rac1-bound lipid kinase complex containing both type I phosphatidylinositol-4-phosphate 5-kinase and diacylglycerol kinase; RhoGDI associates with this lipid kinase complex primarily via its interaction with Rac; the Rac C-terminus is necessary and sufficient for binding to both lipid kinases. In vitro binding with chimeric proteins/peptides/truncation mutants, coimmunoprecipitation, liquid chromatography copurification Molecular and cellular biology Medium 9447972
2003 cAMP elevation (forskolin) in renal CD8 cells decreases active GTP-bound RhoA; coimmunoprecipitation shows RhoA-RhoGDI association increases after forskolin treatment; RhoA phosphorylation on serine (known to stabilize inactive RhoA) increases after cAMP stimulation; RhoA inhibition through phosphorylation and association with RhoGDI is required for AQP2 apical membrane insertion. GTP-RhoA pull-down, cell fractionation, coimmunoprecipitation, western blotting Journal of cell science Medium 12640036
2009 DGKzeta produces phosphatidic acid (PA) which activates PAK1; PAK1 then phosphorylates RhoGDI causing dissociation of Rac1-RhoGDI; DGKzeta stably associates with PAK1 and RhoGDI, forming a complex functioning as a Rac1-selective RhoGDI dissociation factor; DGKzeta-deficient fibroblasts show attenuated PAK1 phosphorylation and Rac1-RhoGDI dissociation, with reduced lamellipodia formation and cell migration. DGKzeta-deficient fibroblasts, coimmunoprecipitation, PAK1 phosphorylation assay, Rac1 activity assay, rescue experiments Molecular biology of the cell High 19211846
2010 DGKalpha produces PA upon HGF stimulation, which recruits atypical PKCzeta/iota in complex with RhoGDI and Rac to ruffling membrane sites; DGKalpha-dependent activation of aPKCzeta/iota releases Rac from its inhibitory complex with RhoGDI, enabling Rac activation and membrane ruffle formation required for cell migration. Coimmunoprecipitation, cell fractionation, dominant-negative constructs, siRNA, fluorescence microscopy PNAS Medium 20160093
2011 XIAP interacts with RhoGDI via the XIAP RING domain; XIAP negatively regulates RhoGDI SUMOylation; XIAP deficiency reduces beta-actin polymerization and cytoskeleton formation, and decreases cell migration and invasion. XIAP knockout/knockdown, coimmunoprecipitation, actin polymerization assay, migration/invasion assay The Journal of biological chemistry Medium 21402697
2012 RhoGDI SUMOylation occurs specifically at Lys-138; SUMOylated RhoGDI has higher binding affinity to Rho GTPases compared to unSUMOylated form; SUMOylation at Lys-138 is crucial for inhibiting actin polymerization, cytoskeleton formation, and cancer cell motility; XIAP RING domain inhibits RhoGDI SUMOylation. Site-directed mutagenesis, coimmunoprecipitation, actin polymerization assay, cell motility assay The Journal of biological chemistry Medium 22393046
2013 PKCalpha phosphorylates RhoGDI1 at Ser96 in pancreatic acini upon CCK stimulation, releasing both RhoA and Rac1 from RhoGDI1; overexpression of RhoGDI1 inhibits RhoA activation and CCK-induced apical amylase secretion; inactive Rac1 influences CCK-induced RhoA activation by preventing RhoGDI1 from binding RhoA. Coimmunoprecipitation, mutagenesis (S96A), RhoGDI overexpression, amylase secretion assay PloS one Medium 23776598
2000 GTP exchange on RhoA is sufficient to trigger its translocation from RhoGDI to liposomes in vitro; GTP-bound G14V-RhoA/RhoGDI complex (but not GDP form) microinjected into cells elicits stress fiber and focal adhesion formation; the GTP-triggered membrane translocation of RhoA from RhoGDI is an intrinsic property not requiring other protein factors or membrane receptors. In vitro liposome translocation assay, microinjection into Swiss 3T3 cells, fluorescence microscopy Protein science High 10716190
2008 Dissociation of Rac1(GDP)•RhoGDI complexes requires cooperative action of anionic liposomes containing PtdIns(3,4,5)P3, a Rac GEF (Trio or Tiam1), and GTP; PtdIns(3,4,5)P3 acts through the GEF pleckstrin homology domain; GEF-mediated GDP-to-GTP exchange on Rac1 drives dissociation; anionic phospholipid composition (not neutral) is the key membrane-level determinant. In vitro reconstitution with purified prenylated Rac1•RhoGDI complexes, liposomes of defined composition, NADPH oxidase activation assay The Journal of biological chemistry High 18505730
1997 The Rho insert region (residues 122-134 of Cdc42Hs) is essential for GDI regulation: a Cdc42Hs/Ha-Ras chimera lacking this insert is insensitive to RhoGDI-mediated inhibition of GDP dissociation and GTP hydrolysis, even though RhoGDI can still bind and extract this chimera from membranes. Chimeric protein construction, in vitro GDI activity assay (GDP dissociation, GTP hydrolysis), membrane extraction assay The Journal of biological chemistry High 9334181
1996 RhoGDI inhibits carboxyl methylation of G25K (Cdc42) in a Mg2+/GDP-dependent manner; RhoGDI and G25K form a stable heterodimer in both GDP- and GTPγS-bound states; RhoGDI co-purifies with the soluble form of G25K from brain. Co-purification, in vitro methylation assay, GTPase binding assay Biochemical and biophysical research communications Medium 8240325
2006 Rac1(W56F) specificity-switch mutant binds RhoGDI and sequesters it, preventing RhoGDI from inactivating RhoA; this elevates cellular GTP-RhoA levels; a dominant-negative RhoA rescues Yersinia invasin-promoted uptake blocked by Rac1(W56F), demonstrating that RhoGDI mediates cross-talk between Rac1 and RhoA activities. FRET, coimmunoprecipitation, GTP-RhoA pull-down, genetic epistasis with dominant-negative RhoA The Journal of biological chemistry Medium 17074770
2001 RhoGDI-binding-defective mutant Cdc42Hs(R66E) is prenylated and membrane-associated (Golgi) but cannot cycle to the cytosol; RhoGDI overexpression translocates wild-type Cdc42Hs but not R66E to cytosol; R66E still activates filopodia formation normally, demonstrating that RhoGDI interaction is required for cytosolic cycling of Cdc42Hs but not for membrane targeting or filopodia induction. Mutagenesis, immunofluorescence, differential centrifugation fractionation, RhoGDI overexpression The Biochemical journal Medium 11583574
2014 RhoGDI facilitates geranylgeranyltransferase-I (GGTase-I)-mediated RhoA prenylation by kinetically trapping the prenylated product, thereby increasing the rate of product release and overall catalytic efficiency; no ternary RhoGDI•RhoA•GGTase-I complex is observed, indicating sequential rather than concurrent action. In vitro prenylation assay, fluorescence-based kinetic measurements Biochemical and biophysical research communications Medium 25223799
2009 In RhoGDI-1(-/-) renal mesangial cells, Rac1 specific activity is selectively elevated (with lesser increases in RhoA and Cdc42); total Rho GTPase protein levels are compensatorily decreased; knockout cells show decreased cell spreading, fewer focal contacts, and reduced proliferation/survival. RhoGDI-1(-/-) cell line characterization, GTPase activity assays, immunofluorescence, morphological analysis Cellular signalling Medium 19765647
2024 PKC-mediated phosphorylation of the RhoGDI N-terminus promotes its interaction with the juxtamembrane domain of p75NTR; MAG induces this PKC phosphorylation, which displaces RIP2 from p75NTR, enhancing RhoA activity and causing stunted neurite outgrowth and apoptosis in cerebellar granule neurons; NGF induces RIP2 recruitment to p75NTR, releasing RhoGDI and decreasing RhoA; the NMR solution structure of the RhoGDI N-terminus/p75NTR juxtamembrane domain complex was determined. NMR structure determination, coimmunoprecipitation, mutagenesis, neurite outgrowth assay, cell survival assay EMBO reports High 38253689
2019 Ang II promotes RhoGDI ubiquitination and proteasomal degradation (at 6 and 48 h); ubiquitin and SUMO competitively modify RhoGDI1 and RhoGDI2, reciprocally regulating their stability; AT1 receptor (not AT2) mediates Ang II-dependent RhoGDI stabilization; RNAi of RhoGDI1 or RhoGDI2 blocks Ang II-induced smooth muscle cell proliferation. Coimmunoprecipitation, proteasome inhibitors, RNAi, proliferation assays (BrdU/EdU), in vivo balloon injury model Atherosclerosis Medium 31362179
2025 KLHL23-Cul3 E3 ligase mediates polyubiquitylation and degradation of CDC42•GTP; KLHL23 and RhoGDI compete for the CDC42 switch II region, with KLHL23 selective for CDC42•GTP and RhoGDI selective for CDC42•GDP; KLHL23 depletion induces excessive membrane protrusions and promotes metastasis; CDC42-Y64C germline variant (Takenouchi-Kosaki Syndrome) escapes KLHL23-mediated degradation; FRET assays show KLHL23 and RhoGDI coordinately inactivate CDC42 in spatiotemporal manner. FRET assays, ubiquitylation assays, KLHL23 depletion, mutagenesis, cell migration/metastasis assays Nature chemical biology High 40846997
2000 The specific binding of isoprenoids (geranylgeranyl/farnesyl) to RhoGDI is saturable in the low micromolar range; carboxymethylated derivatives bind significantly better than free acid counterparts; RhoGDI binding is selective for geranylgeranyl/farnesyl groups and does not accommodate other hydrophobic modifications. Quantitative fluorescence binding assay with rhodamine-labeled isoprenoid analogues Biochemistry Medium 10631002
2021 Andes virus nucleocapsid (N) protein binds the C-terminal domain (residues 69-204) of RhoGDI; this sequesters RhoGDI, reducing available RhoGDI to suppress RhoA; N protein also inhibits RhoA binding to S34D phosphomimetic RhoGDI; collectively, ANDV N protein activates RhoA by reducing free RhoGDI and enhancing release of RhoA from PKC-phosphorylated RhoGDI, increasing endothelial permeability. Coimmunoprecipitation, domain mapping with truncation mutants, phosphomimetic mutagenesis, endothelial permeability assays Journal of virology Medium 34133221

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2001 Differential localization of Rho GTPases in live cells: regulation by hypervariable regions and RhoGDI binding. The Journal of cell biology 587 11149925
1999 Overcoming expression and purification problems of RhoGDI using a family of "parallel" expression vectors. Protein expression and purification 586 10024467
2000 Structure of the Rho family GTP-binding protein Cdc42 in complex with the multifunctional regulator RhoGDI. Cell 424 10676816
2003 The p75 receptor acts as a displacement factor that releases Rho from Rho-GDI. Nature neuroscience 383 12692556
2005 RhoGDI: multiple functions in the regulation of Rho family GTPase activities. The Biochemical journal 336 16083425
2002 Integrins regulate GTP-Rac localized effector interactions through dissociation of Rho-GDI. Nature cell biology 282 11862216
2004 Phosphorylation of RhoGDI by Pak1 mediates dissociation of Rac GTPase. Molecular cell 195 15225553
2013 ARHGDIA mutations cause nephrotic syndrome via defective RHO GTPase signaling. The Journal of clinical investigation 177 23867502
1994 Activation of NADPH oxidase involves the dissociation of p21rac from its inhibitory GDP/GTP exchange protein (rhoGDI) followed by its translocation to the plasma membrane. The Biochemical journal 160 8141770
1998 Characterization of a Rac1- and RhoGDI-associated lipid kinase signaling complex. Molecular and cellular biology 135 9447972
2006 Phosphorylation of RhoGDI by Src regulates Rho GTPase binding and cytosol-membrane cycling. Molecular biology of the cell 132 16943322
2011 Protein kinase A governs a RhoA-RhoGDI protrusion-retraction pacemaker in migrating cells. Nature cell biology 129 21572420
2001 Crystal structure of the Rac1-RhoGDI complex involved in nadph oxidase activation. Biochemistry 120 11513578
1993 A novel role for RhoGDI as an inhibitor of GAP proteins. The EMBO journal 114 8491184
2001 Protein crystallization by rational mutagenesis of surface residues: Lys to Ala mutations promote crystallization of RhoGDI. Acta crystallographica. Section D, Biological crystallography 113 11320308
1996 RhoGDI-3 is a new GDP dissociation inhibitor (GDI). Identification of a non-cytosolic GDI protein interacting with the small GTP-binding proteins RhoB and RhoG. The Journal of biological chemistry 109 8939998
2003 cAMP-induced AQP2 translocation is associated with RhoA inhibition through RhoA phosphorylation and interaction with RhoGDI. Journal of cell science 106 12640036
1996 Characterization of the interaction between RhoGDI and Cdc42Hs using fluorescence spectroscopy. The Journal of biological chemistry 106 8626553
1997 A modulator of rho family G proteins, rhoGDI, binds these G proteins via an immunoglobulin-like domain and a flexible N-terminal arm. Structure (London, England : 1993) 96 9195882
2010 RhoGDI signaling provides targets for cancer therapy. European journal of cancer (Oxford, England : 1990) 78 20347589
2013 ARHGDIA: a novel gene implicated in nephrotic syndrome. Journal of medical genetics 73 23434736
2009 Diacylglycerol kinase zeta regulates actin cytoskeleton reorganization through dissociation of Rac1 from RhoGDI. Molecular biology of the cell 73 19211846
2002 The impact of Glu-->Ala and Glu-->Asp mutations on the crystallization properties of RhoGDI: the structure of RhoGDI at 1.3 A resolution. Acta crystallographica. Section D, Biological crystallography 73 12454455
2011 X-linked inhibitor of apoptosis protein (XIAP) mediates cancer cell motility via Rho GDP dissociation inhibitor (RhoGDI)-dependent regulation of the cytoskeleton. The Journal of biological chemistry 72 21402697
2003 RhoGDI is required for Cdc42-mediated cellular transformation. Current biology : CB 70 12956948
2000 GDP dissociation inhibitor D4-GDI (Rho-GDI 2), but not the homologous rho-GDI 1, is cleaved by caspase-3 during drug-induced apoptosis. The Biochemical journal 69 10698706
1999 How RhoGDI binds Rho. Acta crystallographica. Section D, Biological crystallography 69 10489445
2013 Identification of a novel prenyl and palmitoyl modification at the CaaX motif of Cdc42 that regulates RhoGDI binding. Molecular and cellular biology 68 23358418
2007 RhoGDI-1 modulation of the activity of monomeric RhoGTPase RhoA regulates endothelial barrier function in mouse lungs. Circulation research 68 17525371
2012 RhoGDI SUMOylation at Lys-138 increases its binding activity to Rho GTPase and its inhibiting cancer cell motility. The Journal of biological chemistry 63 22393046
2010 Diacylglycerol kinase alpha mediates HGF-induced Rac activation and membrane ruffling by regulating atypical PKC and RhoGDI. Proceedings of the National Academy of Sciences of the United States of America 61 20160093
2001 Interactions between Rho GTPases and Rho GDP dissociation inhibitor (Rho-GDI). Biochimie 59 11368848
2019 Extraction of active RhoGTPases by RhoGDI regulates spatiotemporal patterning of RhoGTPases. eLife 57 31647414
2001 Structure-activity relationships in flexible protein domains: regulation of rho GTPases by RhoGDI and D4 GDI. Journal of molecular biology 57 11114252
2009 Differential phosphorylation of RhoGDI mediates the distinct cycling of Cdc42 and Rac1 to regulate second-phase insulin secretion. The Journal of biological chemistry 55 20028975
1998 Differential properties of D4/LyGDI versus RhoGDI: phosphorylation and rho GTPase selectivity. FEBS letters 54 9490022
1996 Purification and identification of FOAD-II, a cytosolic protein that regulates secretion in streptolysin-O permeabilized mast cells, as a rac/rhoGDI complex. Molecular biology of the cell 52 8868468
2012 Quantitative analysis of prenylated RhoA interaction with its chaperone, RhoGDI. The Journal of biological chemistry 47 22628549
1997 Interaction between Cdc42Hs and RhoGDI is mediated through the Rho insert region. The Journal of biological chemistry 47 9334181
2002 RhoGDI-3 regulates RhoG and targets this protein to the Golgi complex through its unique N-terminal domain. Traffic (Copenhagen, Denmark) 43 11967128
2004 The impact of Lys-->Arg surface mutations on the crystallization of the globular domain of RhoGDI. Acta crystallographica. Section D, Biological crystallography 41 14747703
2016 Interplay between PCBP2 and miRNA modulates ARHGDIA expression and function in glioma migration and invasion. Oncotarget 39 26761212
2010 RhoGDI: A rheostat for the Rho switch. Small GTPases 39 21686121
2008 Dissociation of Rac1(GDP).RhoGDI complexes by the cooperative action of anionic liposomes containing phosphatidylinositol 3,4,5-trisphosphate, Rac guanine nucleotide exchange factor, and GTP. The Journal of biological chemistry 39 18505730
2012 E3 ligase activity of XIAP RING domain is required for XIAP-mediated cancer cell migration, but not for its RhoGDI binding activity. PloS one 38 22532870
2010 Endocytosis and toxicity of clostridial binary toxins depend on a clathrin-independent pathway regulated by Rho-GDI. Cellular microbiology 38 20846184
1996 Rho guanine nucleotide dissociation inhibitor protein (RhoGDI) inhibits exocytosis in mast cells. The EMBO journal 38 8978674
2000 Mapping the binding site for the GTP-binding protein Rac-1 on its inhibitor RhoGDI-1. Structure (London, England : 1993) 35 10673424
2001 Maintenance of CDC42 GDP-bound state by Rho-GDI inhibits MAP kinase activation by the exchange factor Ras-GRF. evidence for Ras-GRF function being inhibited by Cdc42-GDP but unaffected by CDC42-GTP. The Journal of biological chemistry 33 11285260
2004 Analysis of cell-cycle specific localization of the Rdi1p RhoGDI and the structural determinants required for Cdc42p membrane localization and clustering at sites of polarized growth. Current genetics 32 15108020
2019 RhoGDI stability is regulated by SUMOylation and ubiquitination via the AT1 receptor and participates in Ang II-induced smooth muscle proliferation and vascular remodeling. Atherosclerosis 31 31362179
2006 Liposomes comprising anionic but not neutral phospholipids cause dissociation of Rac(1 or 2) x RhoGDI complexes and support amphiphile-independent NADPH oxidase activation by such complexes. The Journal of biological chemistry 30 16702219
2001 RhoGDI-binding-defective mutant of Cdc42Hs targets to membranes and activates filopodia formation but does not cycle with the cytosol of mammalian cells. The Biochemical journal 30 11583574
2006 Disruption of RhoGDI and RhoA regulation by a Rac1 specificity switch mutant. The Journal of biological chemistry 28 17074770
2008 Downregulation of Rho-GDI gamma promotes differentiation of neural stem cells. Molecular and cellular biochemistry 27 18273563
2000 Differential expression and regulation of GTPases (RhoA and Rac2) and GDIs (LyGDI and RhoGDI) in neutrophils from patients with severe congenital neutropenia. Blood 26 10779444
2013 Overexpression of RhoGDI, a novel predictor of distant metastasis, promotes cell proliferation and migration in hepatocellular carcinoma. FEBS letters 25 24374343
2000 Unique in vivo associations with SmgGDS and RhoGDI and different guanine nucleotide exchange activities exhibited by RhoA, dominant negative RhoA(Asn-19), and activated RhoA(Val-14). The Journal of biological chemistry 24 10702222
2014 SKLB-163, a new benzothiazole-2-thiol derivative, exhibits potent anticancer activity by affecting RhoGDI/JNK-1 signaling pathway. Cell death & disease 23 24675461
2014 Comparative proteomic profiling of triple-negative breast cancer reveals that up-regulation of RhoGDI-2 is associated to the inhibition of caspase 3 and caspase 9. Journal of proteomics 23 24768906
2000 The specific binding of small molecule isoprenoids to rhoGDP dissociation inhibitor (rhoGDI). Biochemistry 23 10631002
2008 The RhoGDI-alpha/JNK signaling pathway plays a significant role in mycophenolic acid-induced apoptosis in an insulin-secreting cell line. Cellular signalling 22 19041939
2015 RhoGDI deficiency induces constitutive activation of Rho GTPases and COX-2 pathways in association with breast cancer progression. Oncotarget 20 26416248
2015 In vivo Rac/Rop localization as well as interaction with RhoGAP and RhoGDI in tobacco pollen tubes: analysis by low-level expression of fluorescent fusion proteins and bimolecular fluorescence complementation. The Plant journal : for cell and molecular biology 19 26252733
2013 Cholecystokinin-mediated RhoGDI phosphorylation via PKCα promotes both RhoA and Rac1 signaling. PloS one 19 23776598
2017 Proteome analysis of irradiated endothelial cells reveals persistent alteration in protein degradation and the RhoGDI and NO signalling pathways. International journal of radiation biology 18 28697312
1993 Carboxyl methylation of the low molecular weight GTP-binding protein G25K: regulation of carboxyl methylation by rhoGDI. Biochemical and biophysical research communications 18 8240325
2007 A proteomic analysis of aorta from spontaneously hypertensive rat: RhoGDI alpha upregulation by angiotensin II via AT(1) receptor. European journal of cell biology 17 17963997
2000 Human RhoA/RhoGDI complex expressed in yeast: GTP exchange is sufficient for translocation of RhoA to liposomes. Protein science : a publication of the Protein Society 17 10716190
2016 Downregulation of ARHGDIA contributes to human glioma progression through activation of Rho GTPase signaling pathway. Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine 16 27726098
2006 Phosphorylation of RhoGDI by p21-activated kinase 1. Methods in enzymology 16 16472651
2008 Genetic disruption of calpain correlates with loss of membrane blebbing and differential expression of RhoGDI-1, cofilin and tropomyosin. The Biochemical journal 15 18076376
2000 Cloning of Rac and Rho-GDI from tobacco using an heterologous two-hybrid screen. Biochimie 15 11120351
2009 Morphological and proliferative abnormalities in renal mesangial cells lacking RhoGDI. Cellular signalling 14 19765647
2014 Differentially expressed proteins in ER+ MCF7 and ER- MDA- MB-231 human breast cancer cells by RhoGDI-α silencing and overexpression. Asian Pacific journal of cancer prevention : APJCP 13 24815488
2001 Crystallization and preliminary crystallographic studies of RhoGDI in complex with the radixin FERM domain. Acta crystallographica. Section D, Biological crystallography 12 11375519
2011 The expression of RKIP, RhoGDI, galectin, c-Myc and p53 in gastrointestinal system of Cr(VI)-exposed rats. Journal of applied toxicology : JAT 11 21437922
2001 Structural consequences of site-directed mutagenesis in flexible protein domains: NMR characterization of the L(55,56)S mutant of RhoGDI. European journal of biochemistry 10 11298742
1997 A somatic cell hybrid panel for distal 17q: GDIA1 maps to 17q25.3. Cytogenetics and cell genetics 10 9186513
2018 Antitumor and radiosensitizing effects of SKLB-163, a novel benzothiazole-2-thiol derivative, on nasopharyngeal carcinoma by affecting the RhoGDI/JNK-1 signaling pathway. Radiotherapy and oncology : journal of the European Society for Therapeutic Radiology and Oncology 9 29519627
2008 Role of Rac and Rho-GDI alpha in the frequency-dependent expression of h1-calponin in vascular smooth muscle cells under cyclic mechanical strain. Annals of biomedical engineering 9 18566890
1999 Regulation of Rho protein binding to membranes by rhoGDI: inhibition of releasing activity by physiological ionic conditions. Biochemistry and cell biology = Biochimie et biologie cellulaire 9 10426287
2019 ROCK inhibitors alleviate myofibroblast transdifferentiation and vascular remodeling via decreasing TGFβ1-mediated RhoGDI expression. General physiology and biophysics 8 31219429
2014 RhoGDI facilitates geranylgeranyltransferase-I-mediated RhoA prenylation. Biochemical and biophysical research communications 8 25223799
2012 Structural coupling between the Rho-insert domain of Cdc42 and the geranylgeranyl binding site of RhoGDI. Biochemistry 8 22206343
2005 RhoGDI-3, a promising system to investigate the regulatory function of rhoGDIs: uncoupling of inhibitory and shuttling functions of rhoGDIs. Biochemical Society transactions 8 16042558
2022 Integrative Network Modeling Highlights the Crucial Roles of Rho-GDI Signaling Pathway in the Progression of non-Small Cell Lung Cancer. IEEE journal of biomedical and health informatics 7 35820010
2012 Cellular function of RhoGDI-α mediates the cycling of Rac1 and the regulation of pancreatic beta cell death. Transplantation proceedings 7 22564631
2014 PRMT3: new binding molecule to RhoGDI-α during mycophenolic acid-induced β-cell death. Transplantation proceedings 6 24815167
2024 RhoGDI phosphorylation by PKC promotes its interaction with death receptor p75NTR to gate axon growth and neuron survival. EMBO reports 5 38253689
2022 Calmodulin Domain Protein Kinase PiCDPK1 Regulates Pollen Tube Growth Polarity through Interaction with RhoGDI. Plants (Basel, Switzerland) 5 35161234
2011 Functional analysis of RhoGDI inhibitory activity on vacuole membrane fusion. The Biochemical journal 4 21171963
2022 TGFβ1 induces myofibroblast transdifferentiation via increasing Smad-mediated RhoGDI-RhoGTPase signaling. General physiology and biophysics 3 36454112
2021 ARHGDIA Confers Selective Advantage to Dissociated Human Pluripotent Stem Cells. Stem cells and development 3 34036793
2011 [Expression of RhoGDI alpha in human testes and sperm and its correlation with the success rate of IVF]. Zhonghua nan ke xue = National journal of andrology 3 21548210
2021 Binding of the Andes Virus Nucleocapsid Protein to RhoGDI Induces the Release and Activation of the Permeability Factor RhoA. Journal of virology 2 34133221
2011 Concordance and interaction of guanine nucleotide dissociation inhibitor (RhoGDI) with RhoA in oogenesis and early development of the sea urchin. Development, growth & differentiation 2 21492154
2025 KLHL23 and RhoGDI coordinate CDC42 inactivation ensuring membrane homeostasis. Nature chemical biology 1 40846997
2012 A quantitative fluorometric approach for measuring the interaction of RhoGDI with membranes and Rho GTPases. Methods in molecular biology (Clifton, N.J.) 1 22144271
2022 miR-497 promotes the migration and invasion of human medulloblast Daoy cell line through targeted regulation of ARHGDIA in vitro. Neuroscience letters 0 35066091

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