Affinage

NET1

Neuroepithelial cell-transforming gene 1 protein · UniProt Q7Z628

Length
596 aa
Mass
67.7 kDa
Annotated
2026-06-11
94 papers in source corpus 24 papers cited in narrative 26 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

NET1 (ARHGEF8/Net1A) is a Dbl-family guanine nucleotide exchange factor specific for RhoA whose oncogenic and morphogenetic activities are governed principally by subcellular partitioning rather than intrinsic catalysis (PMID:8649828, PMID:11839749). In quiescent cells NET1 is held inactive by N-terminal nuclear localization signals that sequester it in the nucleus; removal of the N-terminus or forced cytoplasmic relocalization is sufficient to activate RhoA, drive actin stress-fiber formation, and confer transforming activity (PMID:8649828, PMID:11839749). Cytoplasmic/plasma-membrane NET1 activates RhoA to build stress fibers, mature focal adhesions, phosphorylate myosin light chain, and promote motility and invasion, with the Net1A isoform localizing to focal adhesions and interacting with FAK to control adhesion maturation and the amoeboid-to-mesenchymal invasion switch (PMID:23184663, PMID:23689132, PMID:25588829). Nucleocytoplasmic shuttling integrates multiple upstream inputs: JNK1 phosphorylates Net1A at S52 to drive CRM1-dependent nuclear export (PMID:29361525), a Src→Abl1 cascade phosphorylates Y373 downstream of EGF (PMID:37271338), Rac1→PAK1 phosphorylation at S152/S153 is inhibitory (PMID:15684429), Cdk1 phosphorylation during mitosis suppresses RhoA binding (PMID:33465404), and N-terminal acetylation near the second NLS promotes cytoplasmic accumulation (PMID:25588829). NET1 is a TGF-β/Smad transcriptional target, placing it within the TGF-β→Smad→NET1→RhoA axis that controls cytoskeletal remodeling and EMT (PMID:11278519, PMID:21986943, PMID:20547485). NET1 also performs GEF-activity-independent functions: it stabilizes a Dlg1 scaffold that is required for Src activation (PMID:19586902, PMID:40765140), supports faithful chromosome congression and kinetochore-microtubule attachment in mitosis through a catalytically independent role (PMID:23864709), and in the nucleus binds Smad2 to enhance p300 recruitment and Nodal/TGF-β transcription (PMID:28778986). The 2 Å crystal structure of the RhoA–Net1 DH-domain complex defines an interface that short RhoA-derived peptides can disrupt to block nucleotide exchange (PMID:29695506).

Mechanistic history

Synthesis pass · year-by-year structured walk · 21 steps
  1. 1996 High

    Established NET1 as an oncogene and pinpointed its N-terminus as an autoinhibitory regulatory domain, defining the core problem of how a DH-motif GEF is kept off.

    Evidence Expression cDNA cloning with focus formation and nude-mouse tumorigenicity assays of N-terminally truncated NET1

    PMID:8649828

    Open questions at the time
    • Did not identify the GTPase substrate
    • Mechanism of N-terminal autoinhibition unresolved
  2. 2001 High

    Identified NET1 as a RhoA-specific GEF and placed it downstream of TGF-β/Smad signaling driving stress fiber formation, linking the GEF to a defined morphogenetic pathway.

    Evidence Microarray expression analysis, dominant-negative NET1, ROCK inhibitor, and dominant-negative Smad3 in stress-fiber assays

    PMID:11278519

    Open questions at the time
    • Did not address how NET1 localization gates this activity
    • Smad-to-NET1 transcriptional detail not resolved
  3. 2002 High

    Resolved the autoinhibition mechanism as nuclear sequestration via N-terminal NLSs, showing cytoplasmic relocalization alone activates RhoA — establishing localization as the master switch.

    Evidence Subcellular fractionation, NLS mutagenesis, forced cytoplasmic constructs, and RhoA activation assays

    PMID:11839749

    Open questions at the time
    • Upstream signals controlling shuttling unknown
    • Nuclear function of NET1 not yet defined
  4. 2005 High

    Defined the first inhibitory phosphorylation input, a Rac1→PAK1 axis acting at S152/S153 to down-regulate GEF activity, revealing cross-talk that restrains RhoA.

    Evidence In vitro kinase assay, phospho-specific antibody, Ser→Glu phosphomimetics, and stress-fiber readouts

    PMID:15684429

    Open questions at the time
    • Whether phosphorylation acts via localization or catalysis not fully separated
  5. 2007 High

    Identified Dlg-family tumor suppressors as PDZ-motif partners and showed oncogenic NET1 mislocalizes them, connecting NET1's transforming activity to scaffold sequestration.

    Evidence Reciprocal co-IP, immunofluorescence, PDZ-motif deletion, and transformation rescue assays

    PMID:17938206

    Open questions at the time
    • Direct binding affinity and functional consequence of nuclear Dlg relocalization unresolved
  6. 2008 High

    Demonstrated DNA damage dephosphorylates NET1 to activate a nuclear RhoA pool feeding a p38/MK2 survival response, establishing a stress-responsive nuclear GEF function.

    Evidence Dominant-negative NET1, siRNA, phosphorylation-state analysis, and p38/MK2 pathway readouts after CDT or IR

    PMID:18509476

    Open questions at the time
    • Identity of the inhibitory phosphosite and the phosphatase not defined
  7. 2009 High

    Showed Dlg1 binding stabilizes NET1 against proteasomal degradation and is regulated by cell-cell contact, linking adhesion state to NET1 protein levels and RhoA output.

    Evidence Co-IP, PDZ-domain pulldowns, ubiquitylation and proteasome-inhibitor assays, and calcium/TGF-β perturbation

    PMID:19586902

    Open questions at the time
    • E3 ligase mediating NET1 ubiquitylation not identified
  8. 2011 Medium

    Established that an active nuclear NET1 pool maintains nuclear GTP-RhoA, and that with Ect2 it activates RhoB after IR to promote apoptotic signaling — separating nuclear from cytoplasmic Rho regulation.

    Evidence Active-GEF and GTP-RhoA/RhoB pulldowns from fractions, isolated-nucleus IR experiments, and NET1/Ect2 siRNA with JNK/Bim readouts

    PMID:21373644 PMID:21390328

    Open questions at the time
    • RhoB activation lacks catalytic-mutant validation
    • How IR selectively engages the nuclear pool is unclear
  9. 2011 High

    Resolved isoform-specific regulation, showing TGF-β induces Net1A via Smad/MEK-ERK then drives its degradation and miR-24 suppression during EMT, framing NET1 as a dynamically controlled EMT effector.

    Evidence Isoform-specific RT-PCR, siRNA, proteasome and pathway inhibitors, miR-24 luciferase reporter, and invasion assays

    PMID:21986943

    Open questions at the time
    • Why short-term induction and long-term degradation diverge mechanistically not fully resolved
  10. 2012 High

    Showed Rac1 relocalizes Net1A to the membrane, stimulates its activity, and stabilizes it by a non-catalytic mechanism required for cell spreading and focal adhesion maturation, defining Net1A as the adhesion-active isoform.

    Evidence Active Rac1, proteasome inhibition, isoform-specific siRNA, GEF and MLC phosphorylation assays, and spreading assays

    PMID:23184663

    Open questions at the time
    • Molecular basis of Rac1-driven relocalization not defined at this stage
  11. 2012 Medium

    Linked NET1 to innate/adaptive immune signaling by showing it interacts with CARMA1/3 and cooperates with BCL10 to drive NF-κB activation, broadening its functional repertoire.

    Evidence Co-IP/MS, NF-κB reporter assays, and shRNA knockdown

    PMID:22343628

    Open questions at the time
    • No catalytic-mutant or rescue validation
    • Whether RhoA activity is required is untested
  12. 2013 High

    Identified FAK as a Net1A focal-adhesion partner and uncovered a GEF-independent mitotic role in chromosome congression and centrosomal PAK/Aurora A activation, demonstrating catalysis-independent NET1 functions.

    Evidence Co-IP, focal-adhesion immunofluorescence, invasion assays, catalytically inactive mutant rescue, and SAC/centrosome kinase assays

    PMID:23689132 PMID:23864709

    Open questions at the time
    • Direct molecular effector at kinetochores unknown
    • How GEF-dead NET1 supports mitosis mechanistically unresolved
  13. 2015 High

    Defined N-terminal acetylation near the second NLS as a relocalization signal sufficient for RhoA activation, adding a post-translational layer to the localization switch.

    Evidence Acetylation-site mapping, deacetylase inhibition, Arg/Gln substitutions, and RhoA/focal-adhesion rescue in Net1 KO MEFs

    PMID:25588829

    Open questions at the time
    • Acetyltransferase/deacetylase enzymes acting on Net1A not identified
  14. 2016 Medium

    Extended NET1 into Wnt/β-catenin signaling, showing GEF-dependent activation of PAK1 that phosphorylates β-catenin S675 during dorsal axis formation in vivo.

    Evidence Zebrafish loss/gain-of-function, GEF-dead mutants, PAK1 dimer dissociation, and β-catenin S675 phosphorylation assays

    PMID:27910850

    Open questions at the time
    • Single model organism
    • Conservation of the PAK1/β-catenin link in mammalian cells untested
  15. 2017 Medium

    Established a nuclear, GEF-independent transcriptional role: NET1 binds Smad2 and enhances p300 recruitment to potentiate Nodal/TGF-β signaling, distinguishing NET1's nuclear function from its cytoplasmic GEF activity.

    Evidence Co-IP of NET1 with Smad2 and p300, GEF-dead mutants, and zebrafish Nodal reporter and morphogenesis assays

    PMID:28778986

    Open questions at the time
    • Direct DNA/complex architecture not resolved
    • Mammalian validation lacking
  16. 2018 High

    Defined the JNK1→S52→CRM1 axis driving nuclear export, providing the first complete signal-to-localization mechanism for stress/EGF-induced Net1A activation of RhoA and invasion.

    Evidence In vitro kinase assay, S52A/S52E mutants, CRM1 inhibition, and RhoA/MLC/invasion readouts

    PMID:29361525

    Open questions at the time
    • How JNK is engaged by specific upstream receptors not fully mapped
  17. 2018 High

    Provided the structural basis of catalysis by solving the RhoA–Net1 DH-domain complex at 2 Å and demonstrating interface-disrupting peptides, opening a route to targeted inhibition.

    Evidence X-ray crystallography, molecular dynamics, peptide binding, and GDP-exchange assays

    PMID:29695506

    Open questions at the time
    • No full-length autoinhibited structure
    • Peptide IC50 ~100 µM limits direct utility
  18. 2021 High

    Showed Cdk1 phosphorylates NET1 in mitosis to block RhoA binding and restrain cortical F-actin and spindle polarity, integrating cell-cycle control into NET1 regulation.

    Evidence In vitro Cdk1 assay, phospho-specific antibodies, Ala/Asp mutants, RhoA-interaction and spindle-polarity assays

    PMID:33465404

    Open questions at the time
    • Relationship to the GEF-independent mitotic role from earlier work not reconciled
  19. 2023 High

    Identified a Src→Abl1→Y373 phosphorylation route, distinct from the JNK/S52 axis, that drives EGF-induced cytosolic Net1A and RhoA-dependent invasion, revealing parallel relocalization inputs.

    Evidence EGF stimulation, Src/Abl1 inhibitors, in vitro kinase assay, Y373F/Y373D mutants, and RhoA/MLC/invasion readouts

    PMID:37271338

    Open questions at the time
    • How multiple phospho-inputs are integrated combinatorially not defined
  20. 2024 Medium

    Defined a NET1–HACE1–RAC1 pathway in oocyte meiosis where NET1 protects RAC1 to govern spindle assembly and asymmetric division, broadening NET1's substrate/effector relationships beyond RhoA.

    Evidence Oocyte siRNA, spindle/actin imaging, RAC1 western blot, exogenous RAC1 rescue, and mass spectrometry

    PMID:38169395

    Open questions at the time
    • Mechanism by which NET1 shields RAC1 from HACE1 unresolved
    • Single model system
  21. 2025 Medium

    Demonstrated a GEF-independent requirement for cytosolic NET1 and Dlg1 in full Src activation in cancer cells, separating NET1's RhoA-GEF activity from its scaffold-dependent Src-activating function.

    Evidence Endogenous NET1–Src co-IP, Dlg1 siRNA epistasis, cytosol-targeted and catalytically inactive mutants, and Src Y419/invasion readouts

    PMID:40765140

    Open questions at the time
    • Structural basis of the NET1–Dlg1–Src module not defined
    • Single lab

Open questions

Synthesis pass · forward-looking unresolved questions
  • How the many phospho-, acetylation-, mRNA-localization, and scaffold inputs are integrated to produce context-specific RhoA versus GEF-independent outputs in a given cell remains unresolved.
  • No unified quantitative model of competing localization signals
  • Endogenous regulation across tissues incompletely mapped
  • Mendelian disease association not established in the corpus

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0098772 molecular function regulator activity 5 GO:0060089 molecular transducer activity 3 GO:0140110 transcription regulator activity 1
Localization
GO:0005829 cytosol 5 GO:0005634 nucleus 3 GO:0005815 microtubule organizing center 2 GO:0005886 plasma membrane 2
Pathway
R-HSA-162582 Signal Transduction 5 R-HSA-1640170 Cell Cycle 3 R-HSA-1266738 Developmental Biology 2 R-HSA-5357801 Programmed Cell Death 1 R-HSA-74160 Gene expression (Transcription) 1
Partners
CARMA3DLG1FAKPAK1RAC1RHOASMAD2SRC

Evidence

Reading pass · 26 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1996 NET1 was isolated as a novel oncogene encoding a ~54 kDa protein containing the Dbl-Homology (DH) motif; truncation of its N-terminus activated transforming activity, causing NIH3T3 focus formation and tumorigenicity in nude mice, indicating the N-terminus acts as a negative regulatory domain. Expression cDNA cloning, focus formation assay, nude mouse tumorigenicity assay, fluorescence in situ hybridization (chromosomal mapping to 10p15) Oncogene High 8649828
2002 NET1 localizes to the nucleus via two N-terminal nuclear localization signals (NLS); the oncogenic truncated form lacking the N-terminus is cytoplasmic. Forced cytoplasmic localization of wild-type NET1 is sufficient to activate RhoA, demonstrating that nuclear sequestration is the primary mechanism keeping NET1 inactive. The PH domain additionally functions as a nuclear export signal, independently of catalytic activity. Subcellular fractionation, immunofluorescence, NLS mutation analysis, forced cytoplasmic localization constructs, RhoA activation assays The Journal of biological chemistry High 11839749
2001 NET1 is a guanine nucleotide exchange factor specific for RhoA whose activity is required for TGF-β-induced actin stress fiber formation. TGF-β induces NET1 expression via the Smad signaling pathway, and a dominant-negative NET1 (L392E) or RhoA kinase inhibitor Y-27632 blocks TGF-β-dependent stress fiber formation. Microarray gene expression analysis, overexpression of wild-type and dominant-negative NET1, RhoA kinase inhibitor treatment, dominant-negative Smad3 stable cell line, stress fiber assay The Journal of biological chemistry High 11278519
2005 PAK1 phosphorylates NET1 on serines 152, 153, and 538 in vitro and on S152 in cells. Phosphomimetic substitution at S152/S153 (glutamate) down-regulates NET1 GEF activity in vitro and inhibits stress fiber formation in cells. Rac1 stimulates S152 phosphorylation in a PAK1-dependent manner, establishing a Rac1→PAK1→NET1 inhibitory pathway to suppress RhoA. In vitro kinase assay, phospho-specific antibody, Ser→Glu phosphomimetic mutants, actin stress fiber assay, constitutively active PAK1 co-expression The Journal of biological chemistry High 15684429
2007 NET1 interacts through its C-terminal PDZ-binding motif with tumor suppressor proteins of the Dlg family (Dlg1/SAP97, SAP102, PSD95). This interaction promotes translocation of Dlg proteins to nuclear PML-body-associated subdomains. Oncogenic NET1 (cytoplasmic) sequesters Dlg proteins in the cytosol, reducing their tumor-suppressor activity; co-expression of Dlg1 or SAP102 reduces oncogenic NET1 transforming potential. Co-immunoprecipitation, immunofluorescence, transformation assay, deletion mutant analysis (PDZ-binding motif) Molecular and cellular biology High 17938206
2008 DNA damage (cytolethal distending toxin or ionizing radiation) causes dephosphorylation of NET1 at a critical inhibitory site, activating NET1's GEF activity toward nuclear RhoA. NET1-dependent RhoA activation promotes actin stress fiber formation and cell survival via p38 MAPK and its downstream target MK2. Dominant-negative NET1 expression, siRNA knockdown, phosphorylation state analysis, RhoA activation assay, p38/MK2 pathway readout PloS one High 18509476
2009 NET1 interacts directly with the first two PDZ domains of Dlg1 via its C-terminal PDZ-binding motif. Dlg1 interaction protects NET1 from proteasome-mediated degradation, stabilizing it and increasing NET1-stimulated RhoA activation. Cell-cell contact enhances NET1 stability through increased NET1–Dlg1 interaction; disruption of E-cadherin contacts (by calcium removal or TGF-β) reduces NET1–Dlg1 interaction and promotes NET1 ubiquitylation. Co-immunoprecipitation, pulldown with PDZ domain constructs, ubiquitylation assay, proteasome inhibitor treatment, RhoA activation assay, calcium chelation and TGF-β treatment The Journal of biological chemistry High 19586902
2011 Nuclear NET1 exists in an active (GTP-exchange-competent) form. A fraction of RhoA resides in the nucleus in GTP-bound form, and NET1 activates nuclear RhoA. Ionizing radiation specifically activates the nuclear pool of RhoA via NET1 (nuclear-only), while cytoplasmic RhoA activity remains unchanged; this nuclear NET1/RhoA activation occurs even in isolated nuclei. Affinity precipitation of active GEFs from subcellular fractions, pull-down of GTP-RhoA, siRNA knockdown of NET1, ionizing radiation treatment of whole cells and isolated nuclei PloS one High 21390328
2011 The nuclear GEFs NET1 and Ect2 activate RhoB after ionizing radiation or chemotherapy, promoting apoptotic signaling (JNK phosphorylation, Bim induction). Simultaneous siRNA knockdown of NET1 and Ect2 inhibited IR-induced RhoB activation, reduced apoptotic signaling, and protected cells from IR-induced cell death. siRNA knockdown of NET1 and Ect2, RhoB activation assay, JNK phosphorylation and Bim expression readout, cell death assay PloS one Medium 21373644
2011 TGF-β selectively induces the Net1A isoform (Net1 isoform 2) via Smad and MEK/ERK signaling, leading to cytoplasmic Net1A accumulation and RhoA activation. Long-term TGF-β treatment causes Net1 mRNA downregulation and Net1A protein degradation by the proteasome, triggering EMT. miR-24 post-transcriptionally suppresses Net1A expression and is involved in TGF-β-induced EMT and breast cancer cell invasion. isoform-specific RT-PCR, siRNA knockdown, proteasome inhibitor treatment, Smad/MEK pathway inhibitors, miR-24 luciferase reporter assay, migration/invasion assay Oncogene High 21986943
2012 NET1 interacts with CARMA1 and CARMA3 proteins and induces NF-κB activation. NET1 cooperates with BCL10 and CARMA proteins to stimulate NF-κB activity, and shRNA-mediated NET1 knockdown impairs NF-κB activation by stimuli requiring the CARMA-BCL10-MALT1 complex. Co-immunoprecipitation coupled with mass spectrometry, NF-κB reporter assay, shRNA knockdown The Journal of biological chemistry Medium 22343628
2012 Rac1 activation relocates Net1A from the nucleus to the plasma membrane, stimulates Net1A catalytic activity, and protects Net1A from proteasome-mediated degradation. Net1A (but not Net1) is required for cell spreading on collagen, myosin light chain phosphorylation, and focal adhesion maturation. Net1A relocalization by Rac1 does not require Net1A's catalytic activity, PH domain, or C-terminus, demonstrating a non-catalytic mechanism of regulation. Rac1 constitutively active expression, proteasome inhibitor treatment, immunofluorescence, GEF activity assay, MLC phosphorylation assay, isoform-specific siRNA knockdown, cell spreading assay Molecular and cellular biology High 23184663
2013 Net1A isoform localizes to focal adhesions, interacts with focal adhesion kinase (FAK), and is required for FAK activation, focal adhesion maturation, myosin light chain phosphorylation, and trailing edge retraction during migration. Net1A loss shifts cells from amoeboid to mesenchymal invasion, with elevated β1-integrin and MT1-MMP expression. Co-immunoprecipitation of Net1A with FAK, immunofluorescence to focal adhesions, MLC phosphorylation assay, isoform-specific knockdown, Matrigel invasion assay Molecular and cellular biology High 23689132
2013 NET1 overexpression or knockdown causes mitotic defects including chromosome mis-congression and unstable kinetochore-microtubule attachments, activating the spindle assembly checkpoint. These mitotic functions are independent of RhoA or RhoB activation, as catalytically inactive Net1 rescues mitotic phenotypes. NET1 is required for centrosomal activation of PAK and Aurora A kinase. Net1 overexpression and siRNA knockdown, immunofluorescence for chromosome alignment and kinetochore-microtubule attachments, spindle assembly checkpoint assay, catalytically inactive mutant rescue, centrosome Aurora A and PAK activation assay Molecular biology of the cell High 23864709
2015 Net1A contains two NLS sequences in its N-terminus; residues surrounding the second NLS are acetylated. Deacetylase inhibition or active Rac1 promotes Net1A acetylation and cytoplasmic relocalization. Arginine substitution at the N-terminal acetylation sites prevents cytoplasmic accumulation; glutamine substitution (mimicking acetylation) is sufficient for Net1A relocalization, RhoA activation, F-actin accumulation, and focal adhesion maturation. Acetylation site mapping, deacetylase inhibitor treatment, arginine/glutamine substitution mutants, immunofluorescence, RhoA activation assay, focal adhesion assay, rescue in Net1 KO MEFs Journal of cell science High 25588829
2016 Zebrafish net1 GEF activity is required for Wnt/β-catenin signaling activation during embryonic dorsal axis formation. Net1 dissociates and activates PAK1 dimers; PAK1 then phosphorylates β-catenin on S675, promoting transcription of Wnt target genes. Loss- and gain-of-function in zebrafish embryos, GEF-dead mutant analysis, β-catenin S675 phosphorylation assay, PAK1 dimer dissociation assay Cell research Medium 27910850
2017 Nuclear NET1 interacts with Smad2 in a GEF-activity-independent manner and promotes Smad2 activation by enhancing recruitment of the co-activator p300 to the transcriptional complex, facilitating Nodal signal transduction and mesendoderm formation in zebrafish. Co-immunoprecipitation of Net1 with Smad2 and p300, GEF-dead mutant analysis, zebrafish loss- and gain-of-function experiments, reporter assays for Nodal signaling Journal of cell science Medium 28778986
2018 JNK pathway activation is required and sufficient for Net1A cytosolic relocalization. JNK1 directly phosphorylates Net1A on S52; alanine substitution at S52 prevents relocalization by EGF or JNK activation, while glutamic acid substitution (phosphomimetic) causes spontaneous cytosolic accumulation, elevated RhoA signaling, MLC2 phosphorylation, F-actin accumulation, cell motility, and Matrigel invasion. CRM1 mediates nuclear export of Net1A downstream of JNK. MAPK pathway inhibitors, constitutively active JNK expression, S52A/S52E mutagenesis, CRM1 inhibition (leptomycin B), in vitro kinase assay, immunofluorescence, RhoA activation assay, MLC phosphorylation, invasion assay Journal of cell science High 29361525
2018 Crystal structure of RhoA/Net1 DH-domain heterodimer solved at 2 Å resolution. Structural and molecular dynamics analysis defined the RhoA–Net1 interaction interface. Short RhoA-derived peptides (e.g., EVKHF, residues 102-106) targeting this interface can disrupt the RhoA/Net1 interaction and reduce GEF-catalyzed nucleotide exchange (IC50 ~100 µM). X-ray crystallography (2 Å), molecular dynamics simulation, peptide-based pulldown/binding assay, GDP exchange assay The Journal of biological chemistry High 29695506
2021 Cdk1 phosphorylates NET1 on multiple sites in its N-terminal regulatory domain and C-terminus during mitosis. Substitution of the major Cdk1 phosphorylation sites with acidic residues inhibits NET1's interaction with RhoA; Cdk1 inhibition increases NET1 activity, promotes its plasma membrane localization, and stimulates cortical F-actin accumulation. Acidic substitution of Cdk1 sites reduces Net1-overexpression-induced spindle polarity defects. In vitro Cdk1 kinase assay, phospho-specific antibody generation, Ala/Asp substitution mutants, RhoA interaction assay, immunofluorescence for plasma membrane localization and F-actin, spindle polarity analysis, Cdk1 inhibitor treatment Cellular signalling High 33465404
2023 EGF activates a Src→Abl1 kinase cascade that phosphorylates Net1A on Y373, promoting Net1A cytosolic localization. Y373F substitution prevents cytosolic accumulation; Y373D (phosphomimetic) is sufficient for cytosolic Net1A localization, RhoA activation, MLC2 phosphorylation, F-actin accumulation, cell motility, and Matrigel invasion. Abl1-driven cytosolic localization requires Y373 but acts independently of the JNK-targeted S52 site. EGF stimulation, Src and Abl1 inhibitors, in vitro kinase assay, Y373F/Y373D mutagenesis, immunofluorescence, RhoA activation assay, MLC phosphorylation, invasion assay, Net1A knockdown rescue The Journal of biological chemistry High 37271338
2024 NET1 is required for spindle assembly and actin dynamics during mouse oocyte meiosis. After GVBD, NET1 relocalizes from nucleus to cytoplasm and accumulates on the meiotic spindle at MI/MII. Net1 depletion causes first polar body extrusion failure and asymmetric division defects. NET1 protects RAC1 from HACE1-mediated degradation; exogenous RAC1 expression rescues Net1 depletion phenotypes, establishing a NET1–HACE1–RAC1 pathway governing meiotic cytoskeletal organization. siRNA knockdown in mouse oocytes, immunostaining/confocal microscopy for spindle and actin, western blot for RAC1 levels, exogenous RAC1 rescue experiment, mass spectrometry Reproductive biology and endocrinology Medium 38169395
2025 NET1 cytosolic localization is required for Src kinase activation in breast cancer cells. Endogenous NET1 and Src interact; NET1 expression is required for full Src activation at Y419. This effect requires NET1 cytoplasmic localization and its interaction with scaffolding protein Dlg1, but NOT Net1 catalytic activity toward RhoA. Dlg1 knockdown prevents Src activation by NET1 and blocks Net1–Src interaction. Co-immunoprecipitation of endogenous NET1 and Src, Dlg1 siRNA knockdown, cytosol-targeted and catalytically inactive NET1 mutants, Src Y419 phosphorylation readout, cell motility assay, Matrigel invasion assay Molecular and cellular biology Medium 40765140
2025 NET1/ARHGEF8 is expressed in vascular smooth muscle cells (VSMCs) and is mechanosensitive. Under physiological cyclic stretch, NET1 localizes to the cytosol and interacts with RhoA. Loss of NET1 blunts stretch-induced MYPT1 phosphorylation and impairs cell adhesion and spreading. Expression of a cytosolic NET1 mutant promotes contractile gene expression and increases cell contractile capacity. Cyclic stretch experiments, immunofluorescence for subcellular localization, MYPT1 phosphorylation assay, cell adhesion/spreading assay, cytosolic NET1 mutant expression, gene expression analysis bioRxivpreprint Medium
2024 Net1 mRNA is specifically localized to dermal-epidermal junction (DEJ) protrusion-like structures in stratified squamous epithelia. This mRNA localization dictates Net1 protein distribution and its RhoA GEF activity at that site. Disrupting Net1 mRNA localization alters DEJ morphology and keratinocyte-matrix connections; altered RhoA activity is sufficient to alter DEJ ultrastructure. In situ hybridization/FISH for mRNA localization in mouse epithelium, mRNA localization disruption experiments, RhoA activity assay, electron microscopy of DEJ ultrastructure bioRxivpreprint Medium
2010 In retinal pigment epithelial cells, Smad3 directly transcriptionally induces NET1 expression in response to TGF-β1. NET1 is necessary for TGF-β1-induced RhoA activation and cytoskeletal reorganization (N-cadherin expression, stress fibers); NET1 interacts with RhoA in the cytoplasm. Dominant-negative Smad3 or constitutively active Smad7 blocked NET1 induction and prevented NET1–RhoA interaction. siRNA knockdown of NET1, dominant-negative Smad3 and constitutively active Smad7 cell lines, Co-immunoprecipitation of NET1 with RhoA, RhoA activation assay, immunofluorescence The Journal of biological chemistry High 20547485

Source papers

Stage 0 corpus · 94 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
1999 Net1, a Sir2-associated nucleolar protein required for rDNA silencing and nucleolar integrity. Cell 337 10219245
2005 Support for association between ADHD and two candidate genes: NET1 and DRD1. American journal of medical genetics. Part B, Neuropsychiatric genetics : the official publication of the International Society of Psychiatric Genetics 159 15717291
2001 Net1 stimulates RNA polymerase I transcription and regulates nucleolar structure independently of controlling mitotic exit. Molecular cell 111 11511359
2011 Differential regulation of the two RhoA-specific GEF isoforms Net1/Net1A by TGF-β and miR-24: role in epithelial-to-mesenchymal transition. Oncogene 95 21986943
2001 The activity of guanine exchange factor NET1 is essential for transforming growth factor-beta-mediated stress fiber formation. The Journal of biological chemistry 93 11278519
2008 NET1-mediated RhoA activation facilitates lysophosphatidic acid-induced cell migration and invasion in gastric cancer. British journal of cancer 87 18827818
2011 The small GTPase RhoA localizes to the nucleus and is activated by Net1 and DNA damage signals. PloS one 85 21390328
1996 Isolation of a novel oncogene, NET1, from neuroepithelioma cells by expression cDNA cloning. Oncogene 79 8649828
2002 The Rho exchange factor Net1 is regulated by nuclear sequestration. The Journal of biological chemistry 67 11839749
2006 Net1 and Myeov: computationally identified mediators of gastric cancer. British journal of cancer 66 16552434
2005 PAK1 negatively regulates the activity of the Rho exchange factor NET1. The Journal of biological chemistry 64 15684429
2001 Characterization of the Net1 cell cycle-dependent regulator of the Cdc14 phosphatase from budding yeast. The Journal of biological chemistry 64 11274204
2002 Cdc5 influences phosphorylation of Net1 and disassembly of the RENT complex. BMC molecular biology 63 11960554
2011 The nuclear guanine nucleotide exchange factors Ect2 and Net1 regulate RhoB-mediated cell death after DNA damage. PloS one 62 21373644
2011 Direct effect of morphine on breast cancer cell function in vitro: role of the NET1 gene. British journal of anaesthesia 61 21857017
2002 Mass spectrometry-based methods for phosphorylation site mapping of hyperphosphorylated proteins applied to Net1, a regulator of exit from mitosis in yeast. Molecular & cellular proteomics : MCP 60 12096118
2008 A bacterial cytotoxin identifies the RhoA exchange factor Net1 as a key effector in the response to DNA damage. PloS one 58 18509476
2002 Budding yeast Cdc5 phosphorylates Net1 and assists Cdc14 release from the nucleolus. Biochemical and biophysical research communications 57 12056824
2014 Propofol and bupivacaine in breast cancer cell function in vitro - role of the NET1 gene. Anticancer research 56 24596379
2013 Regulation of focal adhesion kinase activation, breast cancer cell motility, and amoeboid invasion by the RhoA guanine nucleotide exchange factor Net1. Molecular and cellular biology 53 23689132
2010 Smad3 regulates Rho signaling via NET1 in the transforming growth factor-beta-induced epithelial-mesenchymal transition of human retinal pigment epithelial cells. The Journal of biological chemistry 45 20547485
2007 The nuclear RhoA exchange factor Net1 interacts with proteins of the Dlg family, affects their localization, and influences their tumor suppressor activity. Molecular and cellular biology 42 17938206
2019 LncRNA CTC-497E21.4 promotes the progression of gastric cancer via modulating miR-22/NET1 axis through RhoA signaling pathway. Gastric cancer : official journal of the International Gastric Cancer Association and the Japanese Gastric Cancer Association 41 31451992
2021 Simultaneous binding of Guidance Cues NET1 and RGM blocks extracellular NEO1 signaling. Cell 38 33740419
2009 Coexpression of alpha6beta4 integrin and guanine nucleotide exchange factor Net1 identifies node-positive breast cancer patients at high risk for distant metastasis. Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology 37 19124484
2022 Circ_0000745 promotes acute lymphoblastic leukemia progression through mediating miR-494-3p/NET1 axis. Hematology (Amsterdam, Netherlands) 36 34957935
2015 Acetylation of the RhoA GEF Net1A controls its subcellular localization and activity. Journal of cell science 36 25588829
1993 Assignment of the norepinephrine transporter protein (NET1) locus to chromosome 16. Genomics 35 7905857
2021 Baicalin suppresses lung cancer growth phenotypes via miR-340-5p/NET1 axis. Bioengineered 33 33955315
2016 The guanine nucleotide exchange factor Net1 facilitates the specification of dorsal cell fates in zebrafish embryos by promoting maternal β-catenin activation. Cell research 33 27910850
2008 Expression and clinical significance of NET-1 and PCNA in hepatocellular carcinoma. Medical oncology (Northwood, London, England) 31 18214716
2014 miR-22 regulates expression of oncogenic neuro-epithelial transforming gene 1, NET1. The FEBS journal 29 25041463
2012 Rac1 controls the subcellular localization of the Rho guanine nucleotide exchange factor Net1A to regulate focal adhesion formation and cell spreading. Molecular and cellular biology 29 23184663
2018 miR‑22‑3p enhances multi‑chemoresistance by targeting NET1 in bladder cancer cells. Oncology reports 26 29620229
2009 Interaction of the RhoA exchange factor Net1 with discs large homolog 1 protects it from proteasome-mediated degradation and potentiates Net1 activity. The Journal of biological chemistry 26 19586902
2023 AAV-Net1 facilitates the trans-differentiation of supporting cells into hair cells in the murine cochlea. Cellular and molecular life sciences : CMLS 24 36917323
2024 SP1-induced circ_0017552 modulates colon cancer cell proliferation and apoptosis via up-regulation of NET1. Cancer genetics 23 38810361
2013 Inhibition of hepatocellular carcinoma growth and angiogenesis by dual silencing of NET-1 and VEGF. Journal of molecular histology 23 23636606
2018 Contributions of the RhoA guanine nucleotide exchange factor Net1 to polyoma middle T antigen-mediated mammary gland tumorigenesis and metastasis. Breast cancer research : BCR 22 29769144
2011 A functional and transcriptomic analysis of NET1 bioactivity in gastric cancer. BMC cancer 22 21284875
2019 NET1 Enhances Proliferation and Chemoresistance in Acute Lymphoblastic Leukemia Cells. Oncology research 21 31046876
2011 Meiotic nuclear divisions in budding yeast require PP2A(Cdc55)-mediated antagonism of Net1 phosphorylation by Cdk. The Journal of cell biology 19 21690311
2018 Stress-activated MAPKs and CRM1 regulate the subcellular localization of Net1A to control cell motility and invasion. Journal of cell science 18 29361525
2014 The RhoGEF Net1 is required for normal mammary gland development. Molecular endocrinology (Baltimore, Md.) 17 25321414
2012 Neuroepithelial transforming gene 1 (Net1) binds to caspase activation and recruitment domain (CARD)- and membrane-associated guanylate kinase-like domain-containing (CARMA) proteins and regulates nuclear factor κB activation. The Journal of biological chemistry 17 22343628
2013 Rho GTPase-independent regulation of mitotic progression by the RhoGEF Net1. Molecular biology of the cell 16 23864709
2020 Hog1 activation delays mitotic exit via phosphorylation of Net1. Proceedings of the National Academy of Sciences of the United States of America 15 32265285
2021 NET1 promotes HCC growth and metastasis in vitro and in vivo via activating the Akt signaling pathway. Aging 14 33839702
2024 Visible-light-driven net-1,2-hydrogen atom transfer of amidyl radicals to access β-amido ketone derivatives. Chemical science 13 39664809
2019 Evaluation of therapeutic effect of targeting nanobubbles conjugated with NET-1 siRNA by shear wave elastography: an in vivo study of hepatocellular carcinoma bearing mice model. Drug delivery 13 31544556
2012 The effect of NET-1 on the proliferation, migration and endocytosis of the SMMC-7721 HCC cell line. Oncology reports 13 22378020
2019 The C-terminal region of Net1 is an activator of RNA polymerase I transcription with conserved features from yeast to human. PLoS genetics 12 30802237
2019 Cdc14 activation requires coordinated Cdk1-dependent phosphorylation of Net1 and PP2A-Cdc55 at anaphase onset. Cellular and molecular life sciences : CMLS 12 30927017
2014 The possible involvement of genetic variants of NET1 in the etiology of attention-deficit/hyperactivity disorder comorbid with oppositional defiant disorder. Journal of child psychology and psychiatry, and allied disciplines 12 24942521
2011 RhoC and guanine nucleotide exchange factor Net1 in androgen-unresponsive mouse mammary carcinoma SC-4 cells and human prostate cancer after short-term endocrine therapy. The Prostate 12 22095553
2007 The nucleolar Net1/Cfi1-related protein Dnt1 antagonizes the septation initiation network in fission yeast. Molecular biology of the cell 12 17538026
2020 The regulation of Net1/Cdc14 by the Hog1 MAPK upon osmostress unravels a new mechanism regulating mitosis. Cell cycle (Georgetown, Tex.) 11 32794416
2018 Regulation of RhoA activation and cell motility by c-Jun N-terminal kinases and Net1. Small GTPases 11 30332929
2019 Inhibition of NET-1 suppresses proliferation and promotes apoptosis of hepatocellular carcinoma cells by activating the PI3K/AKT signaling pathway. Experimental and therapeutic medicine 10 30867719
2017 Genetic deletion of the Rho GEF Net1 impairs mouse macrophage motility and actin cytoskeletal organization. Small GTPases 10 29173011
2014 Rho GTPase independent regulation of ATM activation and cell survival by the RhoGEF Net1A. Cell cycle (Georgetown, Tex.) 9 25486363
2013 Expression and function of NET-1 in human skin squamous cell carcinoma. Archives of dermatological research 9 24196235
2002 A role for the Saccharomyces cerevisiae RENT complex protein Net1 in HMR silencing. Genetics 9 12196389
2020 MiRNA505/NET1 Axis Acts as a CD8+ T-TIL Regulator in Non-Small Cell Lung Cancer. OncoTargets and therapy 8 33061457
2017 Short interfering RNA targeting Net1 reduces the angiogenesis and tumor growth of in vivo cervical squamous cell carcinoma through VEGF down-regulation. Human pathology 8 28511963
2021 Cdk1 phosphorylation negatively regulates the activity of Net1 towards RhoA during mitosis. Cellular signalling 7 33465404
2021 Quantitative Proteomics Analysis of FFPE Tumor Samples Reveals the Influences of NET-1 siRNA Nanoparticles and Sonodynamic Therapy on Tetraspanin Protein Involved in HCC. Frontiers in molecular biosciences 7 34041269
2020 Propofol suppresses hepatocellular carcinoma by inhibiting NET1 through downregulating ERK/VEGF signaling pathway. Scientific reports 7 32641699
2018 A structural study of the complex between neuroepithelial cell transforming gene 1 (Net1) and RhoA reveals a potential anticancer drug hot spot. The Journal of biological chemistry 7 29695506
2015 Inhibition of skin squamous cell carcinoma proliferation and promote apoptosis by dual silencing of NET-1 and survivin. Oncology reports 7 26080853
2013 Real-world experience of carotid artery stenting in Japan: analysis of 7,134 cases from JR-NET1 and 2 nationwide retrospective multi-center registries. Neurologia medico-chirurgica 7 24305031
2013 Study of RNA Interference Targeting NET-1 Combination with Sorafenib for Hepatocellular Carcinoma Therapy In Vitro and In Vivo. Gastroenterology research and practice 7 24307893
2018 VTIQ evaluates antitumor effects of NET-1 siRNA by UTMD in HCC xenograft models. Oncology letters 6 30127877
2017 A GEF activity-independent function for nuclear Net1 in Nodal signal transduction and mesendoderm formation. Journal of cell science 6 28778986
2024 NET1 is a critical regulator of spindle assembly and actin dynamics in mouse oocytes. Reproductive biology and endocrinology : RB&E 5 38169395
2025 Multiomics analysis reveals the involvement of NET1 in tumour immune regulation and malignant progression. Scientific reports 4 39747410
2006 [Expression of NET-1 gene and protein in hepatocellular carcinoma and related tissues]. Ai zheng = Aizheng = Chinese journal of cancer 4 16536986
2003 NET1 and HFI1 genes of yeast mediate both chromosome maintenance and mitochondrial rho(-) mutagenesis. Yeast (Chichester, England) 4 12898711
2023 Investigation of dynamic solution interactions between NET-1 and UNC-5B by multi-wavelength analytical ultracentrifugation. European biophysics journal : EBJ 3 36939874
2023 Src stimulates Abl-dependent phosphorylation of the guanine exchange factor Net1A to promote its cytosolic localization and cell motility. The Journal of biological chemistry 3 37271338
2018 NET-1 promotes invasion, adhesion and growth of hepatocellular carcinoma by regulating the expression of BAX, caspase 3, caspase 8 and BCL2. Cellular and molecular biology (Noisy-le-Grand, France) 3 30301500
2024 Conditional Overexpression of Net1 Enhances the Trans-Differentiation of Lgr5+ Progenitors into Hair Cells in the Neonatal Mouse Cochlea. Cell proliferation 2 39675772
2009 [Functions of NET-1 gene in skin squamous cell carcinoma cell line (A431): a siRNA study]. Zhonghua bing li xue za zhi = Chinese journal of pathology 2 20078975
2007 [Expression and significance of NET-1 protein in hepatocellular carcinoma]. Zhonghua zhong liu za zhi [Chinese journal of oncology] 2 18478931
2026 Cancer-associated fibroblasts promote osimertinib resistance in non-small cell lung cancer cells via METTL1-mediated NET1 m7G modification. Cell death & disease 1 41723107
2024 Reduced NET1 adversely affects early embryonic development in mice. Theriogenology 1 39426210
2017 Effect of NET-1 siRNA conjugated sub-micron bubble complex combined with low-frequency ultrasound exposure in gene transfection. Oncotarget 1 29423111
2014 Real-world Experience of Carotid Artery Stentingin Japan: Analysis of 7,134 Cases from JR-NET1 and 2Nationwide Retrospective Multi-center Registries. Neurologia medico-chirurgica 1 26236823
2013 Timing is everything: Rac1 controls Net1A localization to regulate cell adhesion. Cell adhesion & migration 1 23792411
2009 [Participation of SRM5/CDC28, SRM8/NET1, and SRM12/HFI1 genes in checkpoint control in yeast Saccharomyces cerevisiae]. Genetika 1 19507699
2025 Net1 Controls Src Activation to Regulate Breast Cancer Cell Motility and Invasion. Molecular and cellular biology 0 40765140
2025 The Role of the RhoA Activating Protein Net1 in Cancer Initiation and Progression. Molecular and cellular biology 0 41449633
2023 Association between NET-1 Level in Placenta from Preeclampsia Pregnancies and Trophoblast Proliferation and Apoptosis: A Correlation Analysis. Alternative therapies in health and medicine 0 37171948
2015 [Influence of ODD diagnosis on the association between NET1 and attention-deficit/hyperactivity disorder]. Zhonghua yi xue za zhi 0 26812975

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