| 2007 |
HEF1/NEDD9 interacts with Aurora A kinase at the basal body of cilia, leading to Aurora A activation, which in turn phosphorylates and activates HDAC6 (a tubulin deacetylase), thereby promoting ciliary disassembly. This constitutes a nonmitotic activity of Aurora A in vertebrates. |
Co-immunoprecipitation, small molecule inhibitors of AurA and HDAC6, loss-of-function and overexpression experiments with defined ciliary resorption phenotype |
Cell |
High |
17604723
|
| 2005 |
HEF1/NEDD9 associates with and activates Aurora A (AurA) kinase at the centrosome; overexpression causes increased centrosome numbers and multipolar spindles. Depletion causes centrosomal splitting, mono-astral spindles, and hyperactivation of Nek2, indicating HEF1 also acts earlier in the cell cycle. |
Co-immunoprecipitation, overexpression and siRNA knockdown with centrosome/spindle phenotype readouts |
Nature cell biology |
High |
16184168
|
| 2006 |
NEDD9 functionally interacts with focal adhesion kinase (FAK) and modulates focal contact formation to enhance melanocyte invasion in vitro and metastasis in vivo. |
Biochemical co-immunoprecipitation, in vitro invasion assays, in vivo metastasis model, focal contact imaging |
Cell |
High |
16814714
|
| 1996 |
Cas-L/NEDD9 (pp105) is tyrosine-phosphorylated upon β1 integrin ligation in T lymphocytes and directly binds both FAK and the adaptor Crk; it also recruits Nck and SHPTP2 upon integrin engagement. |
Immunoprecipitation, in vitro binding assay, peptide sequencing/cloning |
The Journal of experimental medicine |
High |
8879209
|
| 1998 |
HEF1/NEDD9 is processed from full-length p115/p105 forms into a p55 amino-terminal fragment via caspase cleavage specifically at mitosis; p55 localizes to the mitotic spindle. Full-length forms localize to focal adhesions. HEF1 expression is cell-cycle regulated, upregulated upon growth induction. Two-hybrid screening identified the human homolog of G2/M regulator Dim1p as a p55-region interactor. |
Western blotting of synchronized cells, subcellular fractionation/immunofluorescence, yeast two-hybrid screen, caspase inhibitor experiments |
Molecular and cellular biology |
High |
9584194
|
| 2000 |
Smad3 physically interacts with HEF1/NEDD9 and triggers its proteasomal degradation; TGF-β stimulation induces rapid proteasomal degradation of endogenous HEF1 in TGF-β-responsive cells. Elevated HEF1 in turn inhibits TGF-β/Smad3-mediated gene responses, suggesting a negative feedback mechanism. |
Co-immunoprecipitation, overexpression degradation assays, proteasome inhibitor experiments, TGF-β stimulation of endogenous protein |
The EMBO journal |
High |
11118211
|
| 2004 |
Smad3 recruits the APC/C ubiquitin ligase to HEF1/NEDD9 for proteasomal degradation: Smad3 directly contacts APC subunit APC10 via distinct MH2 subdomains, while HEF1 is recognized as a ubiquitination substrate by the CDH1 subunit of APC/C. |
Co-immunoprecipitation, domain mapping, overexpression degradation assays |
BMC cell biology |
Medium |
15144564
|
| 2000 |
HEF1/NEDD9 overexpression induces apoptosis; caspase-dependent cleavage generates 65-, 55-, and 28-kDa fragments during apoptosis. The 28-kDa C-terminal fragment drives apoptosis in a manner dependent on a divergent helix-loop-helix motif and the N-terminal 28 amino acids; it also causes cell rounding. HEF1 overexpression activates JNK, which co-localizes with HEF1. |
Inducible overexpression, caspase inhibitors, deletion/point mutants, immunofluorescence co-localization |
Molecular and cellular biology |
High |
10866674
|
| 2001 |
HEF1/NEDD9 undergoes proteolytic processing specifically in response to cell detachment; integrin receptor ligation and focal adhesion formation prevent this cleavage. A novel focal adhesion targeting domain in the HEF1 C-terminus is required for HEF1-induced cell spreading. |
Cell detachment/attachment assays, integrin blocking experiments, deletion mutant overexpression, immunofluorescence |
Molecular and cellular biology |
High |
11438665
|
| 2002 |
HEF1/NEDD9 inducible expression enhances cell speed and haptotaxis toward fibronectin in a manner partially dependent on ERK and p38 MAPK signaling pathways. HEF1 upregulates mRNAs encoding metalloproteinases, MLCK, p160ROCK, and ErbB2. |
Inducible expression system, cell migration/haptotaxis assays, kinase inhibitors, cDNA expression array |
Journal of cell science |
Medium |
11801728
|
| 2006 |
HEF1/NEDD9 regulates the RhoA activation cycle during mitosis: increased HEF1 sustains RhoA activation causing failure at cytokinesis, while HEF1 depletion reduces RhoA activation causing defects before cleavage furrow formation. HEF1 associates with the RhoA-GEF ECT2. |
siRNA knockdown, overexpression, RhoA activity assays, Co-immunoprecipitation with ECT2, chemical RhoA inhibition rescue |
Molecular biology of the cell |
High |
16394104
|
| 2001 |
FAK regulates β1 integrin-dependent T cell migration through a linear β1 integrin–FAK–HEF1 effector pathway: FAK autophosphorylation and its PR1/HEF1-SH3-binding sites are required for FAK-driven migration and HEF1 tyrosine phosphorylation upon β1 integrin stimulation. |
Overexpression of wild-type and site-specific FAK mutants, siRNA, migration assays, phosphorylation analysis |
European journal of immunology |
Medium |
11465098
|
| 2006 |
HEF1/NEDD9 acts as a necessary and specific downstream effector of FAK in glioblastoma cell migration and invasion: FAK overexpression increases HEF1 phosphorylation; siRNA knockdown of HEF1 (but not p130CAS) inhibits basal and PDGF-stimulated migration and invasion. |
siRNA knockdown, overexpression, FAK mutants (397F), transwell migration/invasion assays, Triton X-100 fractionation |
Oncogene |
Medium |
16288224
|
| 1997 |
HEF1/NEDD9 (p110HEF1) is tyrosine phosphorylated upon β1 integrin ligation in lymphoid cells and associates with CRKL via CRKL's SH2 domain; cell-type-specific differential phosphorylation of HEF1 vs. p120CBL determines different downstream CRKL complexes. |
Co-immunoprecipitation with SH2 domain specificity, anti-integrin stimulation, two cell line comparison |
The Journal of biological chemistry |
Medium |
9162067
|
| 1997 |
HEF1/NEDD9 is tyrosine phosphorylated downstream of BCR/ABL in leukemic cells and forms a complex with Crkl and P190Bcr/Abl; this occurs specifically for HEF1 (not p130Cas), implicating Bcr/Abl in specific interference with β1 integrin signaling via HEF1. |
Immunoprecipitation from transgenic mouse leukemic tissues, phosphotyrosine blotting |
The Journal of biological chemistry |
Medium |
9405482
|
| 1997 |
HEF1/NEDD9 is tyrosine phosphorylated in human B cells upon β1 integrin or BCR ligation in a cytoskeleton-dependent manner; phosphorylated HEF1 associates with RAFTK/Pyk2 and CrkL; cytochalasin B pretreatment blocks both integrin- and BCR-induced HEF1 phosphorylation. |
Co-immunoprecipitation, cytochalasin B treatment, stimulation of primary and cell line B cells |
The Journal of biological chemistry |
Medium |
9020138
|
| 1998 |
Cas-L/NEDD9 is transiently phosphorylated upon TCR/CD3 stimulation; phosphorylated Cas-L binds Crk and C3G. A Cas-L mutant lacking the SH3 domain (FAK-binding site) is phosphorylated by CD3 but not β1 integrin stimulation, indicating FAK-independent TCR signaling through Cas-L. |
CD3 cross-linking, immunoprecipitation, SH3-deletion mutant transfection, phospho-specific blotting |
The Journal of biological chemistry |
Medium |
9497377
|
| 1999 |
The HEF1/NEDD9 C-terminal domain contains a divergent helix-loop-helix (HLH) motif that mediates HEF1 homodimerization and heterodimerization with Id2, E12, and E47 HLH proteins; this HLH is required for HEF1-induced constitutive pseudohyphal growth in yeast. |
Yeast two-hybrid interaction assays, deletion/mutation analysis, yeast pseudohyphal growth assay |
Experimental cell research |
Medium |
10502414
|
| 1999 |
HEF1/NEDD9 (but not p130Cas) is tyrosine phosphorylated downstream of the calcitonin GPCR via Ca2+- and PKC-dependent mechanisms requiring an intact actin cytoskeleton; calcitonin also induces HEF1 association with paxillin and FAK. |
GPCR stimulation, pharmacological inhibitors (BAPTA, calphostin C, cytochalasin D), Co-immunoprecipitation |
The Journal of biological chemistry |
Medium |
10455189
|
| 2000 |
Calcitonin-induced tyrosine phosphorylation of HEF1/NEDD9 requires cell attachment to extracellular matrix via integrin engagement, an intact actin cytoskeleton, and c-Src kinase (specifically SH2-competent Src); overexpression of kinase-dead or SH2-lacking Src blocks HEF1 phosphorylation. Erk1/2 phosphorylation by calcitonin is largely independent of these requirements. |
RGD peptide blocking, poly-D-lysine vs fibronectin adhesion, Src overexpression/mutants, kinase inhibitors |
The Journal of biological chemistry |
Medium |
10954702
|
| 2002 |
TGF-β1 potently induces HEF1/NEDD9 gene transcription in dermal fibroblasts (16-fold protein increase). The p115 HEF1 isoform results from serine/threonine phosphorylation of p105 HEF1. Adhesion-dependent tyrosine phosphorylation of p105HEF1 is mediated by Src kinase (blocked by PP1 inhibitor), while TGF-β1-induced expression is independent of adhesion. |
TGF-β1 stimulation, phosphatase digestion, Src inhibitor PP1, in vitro kinase assay |
The Journal of biological chemistry |
Medium |
12189134
|
| 2005 |
Cell adhesion regulates HEF1/NEDD9 Ser/Thr phosphorylation and proteasomal degradation: actin microfilament disruption activates PP2A which dephosphorylates p115HEF1 to p105HEF1, and the p115 phosphorylated form is preferentially targeted for proteasomal degradation. Adhesion protects HEF1 from degradation. |
Cell detachment, cytoskeletal inhibitors, PP2A inhibitors, proteasome inhibitors, phosphatase assays |
Journal of cell science |
Medium |
16352661
|
| 2009 |
In the MMTV-PyVT mammary tumor model, Nedd9-null background reduces activation of AKT, Src, FAK, and ERK in tumor cells. Cell-derived data show persistently reduced FAK activation and adhesion/migration in Nedd9-/- cells, establishing NEDD9 as a scaffolding protein supporting these pro-oncogenic kinases. |
Genetic knockout mouse model, western blotting of signaling proteins, cell attachment and migration assays |
Cancer research |
High |
19738060
|
| 2013 |
NEDD9 binding to Aurora A kinase stabilizes AURKA protein by limiting CDH1-APC/C ubiquitin ligase binding to AURKA. NEDD9 S296E mutation disrupts binding to AURKA and leads to reduced AURKA protein levels. NEDD9 depletion increases AURKA ubiquitination and proteasomal degradation. |
Co-immunoprecipitation, ubiquitination assays, site-directed mutagenesis (S296E), proteasome inhibitor experiments, shRNA knockdown/re-expression |
Cancer research |
High |
23539442
|
| 2010 |
The WW-HECT E3 ligase Smurf2 physically associates with NEDD9 and is required for NEDD9 protein stability: Smurf2 depletion causes polyubiquitination and proteasomal degradation of NEDD9, while Smurf2 overexpression upregulates NEDD9. The Smurf2/NEDD9 complex is required for Aurora A activation at G2/M and timely mitotic entry. |
Co-immunoprecipitation, siRNA knockdown, ubiquitination assay, Aurora A activity assays, mitotic entry timing |
Cell division |
Medium |
20825672
|
| 2012 |
NEDD9 signaling through integrin β3 leads to elevated phosphorylation of integrin β3, increased Src and FAK activity, and decreased ROCK activity (via Src-dependent phosphorylation of ROCKII Tyr722), driving elongated mesenchymal invasion. Src inhibition by dasatinib switches NEDD9-overexpressing cells from Rac-driven mesenchymal invasion to ROCK-dependent amoeboid invasion. |
Overexpression/knockdown of NEDD9 and integrin β3, Src inhibitor dasatinib, phosphorylation analysis, invasion assays in vitronectin-containing matrix |
Journal of cell science |
Medium |
22328516
|
| 2014 |
NEDD9 regulates cortactin (CTTN) acetylation in an Aurora A kinase (AURKA)/HDAC6-dependent manner: NEDD9 binds to CTTN and its deficiency increases CTTN acetylation and decreases CTTN-F-actin binding, disrupting lamellipodia and migration. A deacetylation-mimicking CTTN-9KR mutant rescues migration defects in NEDD9-depleted cells. |
Co-immunoprecipitation of NEDD9-CTTN, acetylation assays, AURKA/HDAC6 inhibitors, 9KR mutant rescue, xenograft metastasis models |
Molecular cancer research : MCR |
High |
24574519
|
| 2014 |
NEDD9 directly binds to Arf6-GAP ARAP3 and Arf6-effector GGA3, facilitating Arf6 inactivation required for targeting the MMP14/TIMP2 complex to late endosomes. Without NEDD9, MMP14 trafficking is redirected from late endosomes back to the cell surface via Arf6-dependent recycling, preventing TIMP2 disengagement and reducing MMP14 activity and tumor invasion. |
Co-immunoprecipitation of NEDD9-ARAP3/GGA3, Arf6 activity assays, endosomal trafficking imaging, MMP14 activity assays, xenograft models with morpholino antisense |
Oncogene |
High |
25241893
|
| 2013 |
NEDD9 depletion inactivates MMP14 via TIMP2 accumulation at the cell surface, reducing collagenolytic activity of MMP2 and MMP9 and suppressing mesenchymal invasion. Re-expression of NEDD9 restores MMP14 activity. NEDD9 is required for protease-dependent invasion at the primary site but not at the metastatic site. |
shRNA depletion, MMP14 activity assays, TIMP2 immunostaining, in vivo xenograft, re-expression rescue |
Molecular cancer research : MCR |
Medium |
24202705
|
| 2012 |
NEDD9 stabilizes focal adhesions and slows focal adhesion disassembly: NEDD9-/- MEFs show increased paxillin phosphorylation at focal adhesions, increased 2D migration speed, but decreased 3D collagen migration. Loss of NEDD9 suppresses β1 integrin activation and reduces adhesion strength to fibronectin despite upregulated α5β1 expression. |
NEDD9-/- mouse embryo fibroblasts, focal adhesion turnover assays, 2D/3D migration assays, integrin activation assays, adhesion strength measurement |
PloS one |
Medium |
22509381
|
| 2011 |
NEDD9 and BCAR1 signal through SRC to promote E-cadherin removal from the cell membrane and lysosomal degradation, without affecting E-cadherin transcription. Nedd9-/- mammary tumors show enhanced junctional E-cadherin. |
siRNA knockdown, Src inhibitors, membrane fractionation, lysosomal inhibitors, immunofluorescence, Nedd9-/- mouse mammary tumor model |
PloS one |
Medium |
21765937
|
| 2000 |
AND-34 (a GEF) associates with HEF1/NEDD9 via its GEF domain binding to the HEF1 C-terminus; AND-34 overexpression activates Cdc42 (but not Rac, Rho, RalA, or Rap1) in B cells, enhancing PAK1 activity and inhibiting SDF-1α-induced B cell polarization. |
Co-immunoprecipitation, GTPase pulldown assays, PAK1 kinase assay, overexpression |
The Journal of biological chemistry |
Medium |
10692442
|
| 2006 |
Chat-H (hematopoietic isoform of Cas/HEF1-associated signal transducer) associates with HEF1/NEDD9 (CasL) and mediates serine-threonine phosphorylation of CasL; Chat-H localization to the plasma membrane and its binding to CasL are required for T cell migration and chemokine-induced Rap1 activation. |
Lentiviral RNAi, co-immunoprecipitation, phosphorylation analysis, Rap1 activation assay, T cell migration assay |
Immunity |
Medium |
17174122
|
| 2012 |
Abl and Arg tyrosine kinases mediate chemokine-induced tyrosine phosphorylation of HEF1/NEDD9; phosphorylated HEF1 is required for Rap1 GTPase activation, which mediates T cell adhesion and migration. T cells lacking Abl and Arg exhibit defective lymph node homing and impaired migration to inflammation sites. |
Abl/Arg knockout T cells, co-immunoprecipitation, Rap1 activation assay, in vivo homing assay, transwell migration |
Science signaling |
Medium |
22810897
|
| 2012 |
LKB1 negatively regulates NEDD9 transcription by promoting cytosolic translocation of CRTC1 from the nucleus. Ectopic NEDD9 or CRTC1 expression partially reverses the inhibitory function of LKB1 on lung cancer metastasis, establishing a CRTC1-NEDD9 axis downstream of LKB1. |
RNAi silencing in de novo mouse lung tumor models, ectopic expression, subcellular fractionation of CRTC1, metastasis assays |
Cancer research |
Medium |
23074285
|
| 2011 |
HEF1/NEDD9 is a direct transcriptional target of canonical Wnt/β-catenin signaling: ChIP assays and promoter analyses identified three functional TCF-binding sites in the HEF1 promoter responsible for Wnt-3a/β-catenin/Dvl2-driven upregulation; shRNA knockdown of β-catenin suppresses HEF1 expression. |
Chromatin immunoprecipitation (ChIP), promoter luciferase assays, shRNA knockdown, Wnt-3a stimulation |
Oncogene |
Medium |
21317929
|
| 2010 |
VHL inactivation induces HEF1/NEDD9 and Aurora kinase A via stabilization of HIF-1 and HIF-2. HEF1/NEDD9 colocalizes with Aurora kinase A at the centrosome and enhances Aurora A's cilium-destabilizing effect; suppression of this pathway improves primary cilium formation and reduces motility in VHL-defective renal cancer cells. |
siRNA knockdown, immunofluorescence co-localization, VHL re-expression, HIF stabilization, cilia formation assay |
Journal of the American Society of Nephrology : JASN |
Medium |
20864688
|
| 2010 |
HEF1/NEDD9 is required for VEGF-mediated head and neck cancer cell migration and invasion; VEGF promotes HEF1-dependent invadopodia formation, and HEF1 co-localizes with MT1-MMP at invadopodia. |
siRNA knockdown and overexpression, phosphotyrosine proteomics, invadopodia assays, co-immunolocalization |
Oncogene |
Medium |
20498643
|
| 2018 |
Oxidation of Cys18 in the SMAD3 docking region of NEDD9 impairs SMAD3-NEDD9 protein-protein interaction in vitro (demonstrated by microscale thermophoresis). Aldosterone-induced oxidant stress in pulmonary artery endothelial cells reproduces this effect, resulting in impaired NEDD9 proteolytic degradation, increased NEDD9 complex formation with NKX2-5, and increased NKX2-5 binding to the COL3A1 promoter to upregulate collagen III. NEDD9 ablation prevents fibrotic vascular remodeling in animal PAH models. |
Microscale thermophoresis (in vitro protein interaction), ALDO-induced oxidant stress, Co-immunoprecipitation (NEDD9-NKX2-5), ChIP (NKX2-5 on COL3A1), atomic force microscopy, NEDD9 knockout animal model |
Science translational medicine |
High |
29899023
|
| 2021 |
NEDD9 mediates platelet adhesion to pulmonary artery endothelial cells via a NEDD9-P-selectin interaction; under hypoxia, HIF-1α-dependent NEDD9 upregulation increases surface NEDD9. Anti-NEDD9 antibody targeting the NEDD9-P-selectin interaction inhibits platelet-endothelial adhesion in vitro and reduces pulmonary thromboembolic remodeling in vivo in NEDD9-/- mice. |
Network medicine/proteomics identification, microscale thermophoresis, NEDD9-/- mice, anti-NEDD9 antibody functional assay, ex vivo CTEPH patient cells |
American journal of respiratory and critical care medicine |
Medium |
33523764
|
| 2016 |
Cas-L/NEDD9 is phosphorylated at TCR microclusters in an actin polymerization-dependent manner and is required for transport of TCR microclusters to the center of the immunological synapse; Cas-L participates in a positive feedback loop amplifying Ca2+ signaling, inside-out integrin activation, and actomyosin contraction, acting as a mechanical transducer linking TCR to actin. |
siRNA knockdown, super-resolution imaging of TCR microclusters, Ca2+ imaging, integrin activation assay, traction force microscopy |
Immunology and cell biology |
Medium |
27359298
|
| 2009 |
Loss of Nedd9 in chick neural crest cells perturbs cell spreading, reduces focal complex density and actin filaments, and causes a graded reduction in migratory distance in vivo. Retinoic acid regulates Nedd9 expression in neural crest cells. |
siRNA knockdown and overexpression in chick neural crest, in ovo electroporation, immunofluorescence of focal complexes and actin |
Neuroscience |
Medium |
19464348
|
| 2009 |
AhR/dioxin activation transcriptionally induces Nedd9/HEF1 via two xenobiotic response elements (XREs) in its promoter; RNAi knockdown of Nedd9 blocks dioxin-induced changes in adhesion, cytoskeleton reorganization, increased cell migration, E-cadherin repression, and JNK activation. |
XRE reporter assay, RNAi knockdown, AhR ligand stimulation, migration/adhesion assays |
Oncogene |
Medium |
19648964
|
| 2008 |
NEDD9 is directly regulated by all-trans retinoic acid (atRA) through a complex retinoic acid response element (RARE) located at -475 to -445 in the NEDD9 proximal promoter; RAR and RXR are physically bound to this RARE in cells, as shown by ChIP. |
Promoter luciferase assays with RARE mutations, EMSA, ChIP for RAR/RXR binding |
Archives of biochemistry and biophysics |
Medium |
18585997
|
| 2014 |
Pkd1-/-;Nedd9-/- mice show striking morphological ciliary defects with specific loss of ciliary localization of adenylyl cyclase III, and have ciliary resorption defects compatible with failure of Aurora A activation. Cystogenesis is strongly promoted in the double-mutant, indicating Nedd9 acts as a modifier of ADPKD via Aurora A-dependent ciliary maintenance. |
Compound Pkd1/Nedd9 genetic knockout mice, immunofluorescence of cilia, Aurora A activity analysis, cystogenesis quantification, calcium response assays |
Proceedings of the National Academy of Sciences of the United States of America |
Medium |
25139996
|
| 2023 |
Pan-HDAC inhibitors enhance H3K9 acetylation at the NEDD9 gene promoter via inhibition of HDAC4 activity, increasing NEDD9 expression and subsequent FAK phosphorylation activation, promoting breast cancer metastasis. FAK inhibitors can reverse this metastasis. |
ChIP for H3K9 acetylation at NEDD9 promoter, HDAC4 inhibition, FAK phosphorylation assays, invasion assays, preclinical metastasis models |
Signal transduction and targeted therapy |
Medium |
36604412
|
| 2018 |
NEDD9 stimulates MMP9 secretion and invadopodia formation through its substrate domain (SD) tyrosine phosphorylation and SH3 domain; mutation of all 13 YxxP motif tyrosines plus Y629 (F14NEDD9) eliminates tyrosine phosphorylation, MMP9 secretion, and invadopodia. MICAL1 silencing also reduces MMP9 secretion, suggesting NEDD9-MICAL1 cooperation. |
Stable expression of NEDD9 domain mutants, MMP9 secretion assays, invadopodia formation assays, MICAL1 shRNA |
Oncotarget |
Medium |
29876004
|
| 2005 |
HTLV-I Tax physically associates with Cas-L/NEDD9, binding through the serine-rich region of Cas-L; Tax increases Cas-L expression and tyrosine phosphorylation. Exogenous Cas-L inhibits Tax-mediated NF-κB transactivation but not Tax-independent NF-κB activation. |
Yeast two-hybrid screen, co-immunoprecipitation, NF-κB reporter assay, co-localization |
Oncogene |
Medium |
15592516
|