Affinage

HECW2

E3 ubiquitin-protein ligase HECW2 · UniProt Q9P2P5

Length
1572 aa
Mass
175.8 kDa
Annotated
2026-06-10
31 papers in source corpus 10 papers cited in narrative 10 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 5/5 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

HECW2 (NEDL2) is a HECT-domain E3 ligase that uses its WW domains to engage PY- and PIP-motif-bearing substrates and exerts dual catalytic outputs—classical ubiquitin transfer and an intrinsic Nedd8 ligase activity dependent on catalytic cysteine 1341 (PMID:27119228). Its ubiquitination outcomes are substrate-dependent: it ubiquitinates p73 yet stabilizes it to enhance p73-driven transcription (PMID:12890487), deposits non-degradative K63-linked chains on AMOTL1 to reinforce endothelial cell-cell junctions and retain YAP in the cytoplasm, restraining angiogenic sprouting (PMID:27498087), and conversely directs proteasomal degradation of PCNA, lamin B1, lamin A, and the heterochromatin proteins HP1α and HP1β (PMID:29753763, PMID:30208514). Beyond catalysis, HECW2 functions as a scaffold that assembles a SHC/Grb2/PI3K/PDK1/Akt complex downstream of GDNF/Ret, and its Nedd8 ligase activity—not its ubiquitin ligase activity—is required for GDNF-stimulated Akt signaling and for enteric nervous system and kidney development in vivo (PMID:25555806, PMID:27119228). HECW2 is itself cell-cycle-regulated: APC/C-Cdh1 recognizes a destruction box and targets it for degradation at mitotic exit, and HECW2 associates with mitotic spindles and influences the metaphase-to-anaphase transition (PMID:24163370). In disease contexts, HECW2-mediated lamin B1 degradation activates AKT/mTOR to promote colorectal cancer progression and chemoresistance (PMID:37781079), and HECW2 ubiquitination of the m6A demethylase ALKBH5 drives LDHA-dependent glycolysis in hemangioma endothelial cells (PMID:40520873).

Mechanistic history

Synthesis pass · year-by-year structured walk · 9 steps
  1. 2003 Medium

    Established the founding paradigm that HECW2 is an E3 ligase whose ubiquitination of a substrate can stabilize rather than degrade it, coupling HECW2 to p73 transcriptional output.

    Evidence Reciprocal Co-IP, in vitro pull-down and ubiquitination assays, stability/decay measurements, and transcriptional reporter assays for the HECW2–p73 interaction

    PMID:12890487

    Open questions at the time
    • Chain linkage type and the mechanism by which ubiquitination stabilizes p73 not resolved
    • No in vivo confirmation of the HECW2–p73 axis
  2. 2013 High

    Placed HECW2 in the cell cycle by showing it is a degradation substrate of APC/C-Cdh1 via a destruction box and that its levels and spindle association couple it to mitotic progression.

    Evidence MS-identified interaction, reciprocal Co-IP in vivo and in vitro, destruction-box mutant analysis, and knockdown/overexpression mitotic phenotypes

    PMID:24163370

    Open questions at the time
    • Mitotic substrates of HECW2 not identified
    • Mechanism linking HECW2 to metaphase delay and chromosomal lagging unresolved
  3. 2014 High

    Demonstrated a required physiological role for HECW2 by showing knockout mice develop intestinal aganglionosis, tying HECW2 to GDNF/Ret/Akt-driven enteric neural precursor proliferation.

    Evidence HECW2 knockout mouse with histological/functional intestinal readouts and Akt phosphorylation analysis

    PMID:25555806

    Open questions at the time
    • Molecular mechanism of pathway engagement not defined in this study
    • Catalytic activity requirement not yet dissected
  4. 2016 High

    Resolved the GDNF mechanism as scaffold assembly and uncovered an intrinsic Nedd8 ligase activity (Cys1341), showing neddylation—not ubiquitination—drives Akt activation and development.

    Evidence Scaffold co-IP assembly assays, Nedd8 ligase assay, C1341 site-directed mutagenesis, and double-knockout mouse ENS/kidney phenotypes

    PMID:27119228

    Open questions at the time
    • Neddylation target(s) within the GDNF complex not identified
    • How a single protein toggles between ubiquitin and Nedd8 ligase outputs unknown
  5. 2016 High

    Extended the stabilizing-ubiquitination paradigm to endothelial biology, showing K63-linked ubiquitination of AMOTL1 maintains junctions and restrains YAP nuclear translocation and angiogenesis.

    Evidence Reciprocal Co-IP, K63-linkage-specific ubiquitination assay, siRNA knockdown, YAP localization imaging, sprouting assay, and ANG-2 epistasis

    PMID:27498087

    Open questions at the time
    • Direct mechanistic link between AMOTL1 K63 chains and YAP sequestration not fully mapped
    • Single-lab evidence
  6. 2018 Medium

    Identified degradative substrates of HECW2—PCNA (via a PIP motif), lamin B1, lamin A, HP1α and HP1β—broadening its role to nuclear architecture and heterochromatin protein turnover.

    Evidence Co-IP, PIP-motif-dependent interaction, ubiquitination assays, proteasome inhibitor rescue, lamin A mutant comparison, and HP1-isoform knockdown westerns

    PMID:29753763 PMID:30208514

    Open questions at the time
    • Cellular consequences of substrate degradation not established
    • Reduced binding to laminopathy mutants not linked to disease phenotype
  7. 2021 Medium

    Localized HECW2 to germinal-vesicle and pronuclear nuclei and implicated it functionally in sperm DNA decondensation and male pronuclear formation, with distinct tissue-specific isoforms.

    Evidence Western-blot isoform analysis, immunofluorescence across developmental stages, and anti-NEDL2 antibody microinjection in porcine IVF

    PMID:33030211

    Open questions at the time
    • Substrate mediating pronuclear formation unknown
    • Functional roles of the distinct isoforms not characterized
  8. 2023 Medium

    Connected HECW2 substrate degradation to oncogenic signaling, showing lamin B1 degradation activates AKT/mTOR to drive colorectal cancer progression and chemoresistance.

    Evidence HECW2 knockdown/overexpression in CRC cells, lamin B1 ubiquitination assay, AKT/mTOR western blots, and proliferation/chemoresistance assays

    PMID:37781079

    Open questions at the time
    • Mechanistic link between lamin B1 loss and AKT/mTOR activation not detailed
    • Single-lab evidence
  9. 2025 Medium

    Linked HECW2 to RNA metabolism and tumor metabolism by showing it ubiquitinates the m6A demethylase ALKBH5, which enhances LDHA expression and glycolysis in hemangioma endothelium.

    Evidence HECW2 overexpression/knockdown, ALKBH5 ubiquitination assay, m6A demethylation readout on LDHA mRNA, glycolysis assays, and xenograft model

    PMID:40520873

    Open questions at the time
    • Whether ALKBH5 ubiquitination is degradative or activating not clarified
    • Single-lab evidence

Open questions

Synthesis pass · forward-looking unresolved questions
  • How HECW2 selects between stabilizing (non-degradative/K63) and degradative ubiquitination, and between ubiquitin and Nedd8 ligase outputs, across its diverse substrates remains unresolved.
  • No structural model of HECW2 catalytic switching
  • Substrate-specific cofactors or regulatory inputs unknown
  • Neddylation substrates largely unidentified

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0140096 catalytic activity, acting on a protein 6 GO:0098772 molecular function regulator activity 2 GO:0016874 ligase activity 1 GO:0060090 molecular adaptor activity 1
Localization
GO:0005634 nucleus 1
Pathway
R-HSA-162582 Signal Transduction 4 R-HSA-392499 Metabolism of proteins 4 R-HSA-1266738 Developmental Biology 2 R-HSA-1640170 Cell Cycle 1
Partners
ALKBH5AMOTL1CBX5FZR1LMNALMNB1PCNATP73
Complex memberships
GDNF/Ret/PI3K/Akt signaling complex

Evidence

Reading pass · 10 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2003 HECW2 (NEDL2) physically interacts with p73 via its WW domains recognizing PY motifs on p73, ubiquitinates p73, and paradoxically stabilizes rather than degrades it, thereby enhancing p73-dependent transcriptional activation. Reciprocal co-immunoprecipitation, in vitro pull-down assay, in vitro ubiquitination assay, p73 stability/decay measurements, transcriptional reporter assay Biochemical and biophysical research communications Medium 12890487
2013 HECW2 (NEDL2) is a substrate of APC/C-Cdh1 ubiquitin ligase; Cdh1 recognizes a destruction box (R740GSL743) on NEDL2 and targets it for proteasomal degradation during mitotic exit. NEDL2 associates with mitotic spindles, its protein level peaks in mitosis, and its depletion prolongs metaphase while overexpression induces chromosomal lagging. Co-immunoprecipitation, mass spectrometry, in vitro interaction assay, overexpression and knockdown experiments with mitotic phenotype readouts, destruction-box mutant analysis The Journal of biological chemistry High 24163370
2014 HECW2 (NEDL2) positively regulates enteric neural precursor proliferation through the GDNF/Ret/Akt signaling pathway; NEDL2-deficient mice display intestinal aganglionosis and postnatal lethality, establishing a required in vivo role. NEDL2 knockout mouse model with histological and functional readout (intestinal aganglionosis, bowel motility), signaling pathway analysis (Akt phosphorylation) Cellular signalling High 25555806
2016 HECW2 (NEDL2) acts as a scaffold to recruit SHC, Grb2, PI3K (p110/p85), PDK1, and Akt, promoting GDNF-stimulated Akt signaling. Additionally, NEDL2 harbors intrinsic Nedd8 ligase activity with cysteine 1341 as the catalytic site, and Nedd8 ligase—but not ubiquitin ligase—activity is required for GDNF-stimulated Akt activation and ENS/kidney development. Biochemical co-immunoprecipitation/scaffold assembly assay, Nedd8 ligase activity assay, site-directed mutagenesis (C1341), double knockout mouse phenotype, kidney development analysis Oncotarget High 27119228
2016 HECW2 physically interacts with AMOTL1 and stabilizes it via K63-linked ubiquitination. HECW2 depletion in human endothelial cells decreases AMOTL1 stability, loosens cell-to-cell junctions, relocalizes YAP from cytoplasm to nucleus, and increases angiogenic sprouting—an effect blocked by ANG-2 depletion. Co-immunoprecipitation, ubiquitination assay specifying K63-linkage, siRNA knockdown, YAP subcellular localization (imaging), sprouting angiogenesis assay, epistasis (ANG-2 co-depletion) Cellular signalling High 27498087
2018 HECW2 interacts with PCNA via a canonical PIP motif and with lamin B1, ubiquitinates both, and targets them for proteasomal degradation. HECW2 also interacts with wild-type lamin A and ubiquitinates it, but this interaction is reduced with laminopathy-causing lamin A mutants (G232E, Q294P). Co-immunoprecipitation, PIP-motif-dependent interaction assay, ubiquitination assay, proteasome inhibitor rescue, lamin A mutant cell lines Biochimica et biophysica acta. Molecular cell research Medium 29753763
2018 HECW2 interacts with HP1α and HP1β (but not HP1γ), ubiquitinates them, and targets them for proteasomal degradation; HECW2 downregulation increases steady-state levels of HP1α and HP1β. Co-immunoprecipitation, ubiquitination assay, siRNA knockdown with western blot quantification of HP1 isoforms Biochemical and biophysical research communications Medium 30208514
2021 HECW2 (NEDL2) localizes to nuclear foci in germinal vesicles of immature oocytes, to pronuclei of zygotes, and to nuclei of blastomeres. Multiple protein isoforms exist with distinct tissue distributions (52 kDa in germ cells/embryos; 91/136/155 kDa in somatic cells). Microinjection of anti-NEDL2 antibody into porcine oocytes inhibited sperm DNA decondensation and male pronuclear formation without affecting fertilization rate or blastocyst formation. Western blot isoform analysis, immunofluorescence localization across developmental stages, antibody microinjection functional assay in porcine IVF Biology of reproduction Medium 33030211
2023 HECW2 promotes colorectal cancer progression and chemoresistance by mediating ubiquitin-proteasome degradation of lamin B1, which activates the AKT/mTOR signaling pathway. HECW2 knockdown and overexpression in CRC cells, ubiquitination assay of lamin B1, AKT/mTOR pathway analysis by western blot, functional assays (proliferation, chemoresistance) Journal of Cancer Medium 37781079
2025 HECW2 regulates ubiquitination of ALKBH5 (an m6A RNA demethylase), which subsequently enhances LDHA expression through ALKBH5-mediated m6A demethylation of LDHA mRNA, promoting glycolysis in hemangioma endothelial cells. HECW2 overexpression/knockdown, ubiquitination assay of ALKBH5, m6A demethylation readout on LDHA mRNA, functional glycolysis assays (glucose uptake, lactate, ATP), in vivo xenograft model American journal of cancer research Medium 40520873

Source papers

Stage 0 corpus · 31 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2003 A novel HECT-type E3 ubiquitin ligase, NEDL2, stabilizes p73 and enhances its transcriptional activity. Biochemical and biophysical research communications 86 12890487
2018 Engagement of circular RNA HECW2 in the nonautophagic role of ATG5 implicated in the endothelial-mesenchymal transition. Autophagy 83 29260931
2016 Mutations in HECW2 are associated with intellectual disability and epilepsy. Journal of medical genetics 59 27334371
2016 The endothelial E3 ligase HECW2 promotes endothelial cell junctions by increasing AMOTL1 protein stability via K63-linked ubiquitination. Cellular signalling 44 27498087
2013 The HECT type ubiquitin ligase NEDL2 is degraded by anaphase-promoting complex/cyclosome (APC/C)-Cdh1, and its tight regulation maintains the metaphase to anaphase transition. The Journal of biological chemistry 40 24163370
2018 E3 ubiquitin ligase HECW2 targets PCNA and lamin B1. Biochimica et biophysica acta. Molecular cell research 35 29753763
2020 Circ_HECW2 functions as a miR-30e-5p sponge to regulate LPS-induced endothelial-mesenchymal transition by mediating NEGR1 expression. Brain research 23 32916175
2010 Knobbed acrosome defect is associated with a region containing the genes STK17b and HECW2 on porcine chromosome 15. BMC genomics 23 21143916
2014 NEDL2 is an essential regulator of enteric neural development and GDNF/Ret signaling. Cellular signalling 18 25555806
2016 NEDL2 regulates enteric nervous system and kidney development in its Nedd8 ligase activity-dependent manner. Oncotarget 14 27119228
2021 NEDD4-like ubiquitin ligase 2 protein (NEDL2) in porcine spermatozoa, oocytes, and preimplantation embryos and its role in oocyte fertilization†. Biology of reproduction 13 33030211
2025 Circ_HECW2 regulates ox-LDL-induced dysfunction of cardiovascular endothelial cells by miR-942-5p/TLR4 axis. Clinical hemorheology and microcirculation 12 36213989
2018 E3 ubiquitin ligase HECW2 mediates the proteasomal degradation of HP1 isoforms. Biochemical and biophysical research communications 12 30208514
2021 HECW2-related disorder in four Japanese patients. American journal of medical genetics. Part A 11 34047014
2018 Rett-like features and cortical visual impairment in a Japanese patient with HECW2 mutation. Brain & development 10 29395664
2021 Circ_HECW2 regulates LPS-induced apoptosis of chondrocytes via miR-93 methylation. Immunity, inflammation and disease 9 34076365
2020 Association of HECW2 variants with developmental and epileptic encephalopathy and knockdown of zebrafish hecw2a. American journal of medical genetics. Part A 9 33205896
2022 A homozygous nonsense HECW2 variant is associated with neurodevelopmental delay and intellectual disability. European journal of medical genetics 7 35487419
2023 HECW2 promotes the progression and chemoresistance of colorectal cancer via AKT/mTOR signaling activation by mediating the ubiquitin-proteasome degradation of lamin B1. Journal of Cancer 6 37781079
2021 A novel likely pathogenic heterozygous HECW2 missense variant in a family with variable expressivity of neurodevelopmental delay, hypotonia, and epileptiform EEG patterns. American journal of medical genetics. Part A 6 34327820
2024 Mechanism of Circ_HECW2 regulating osteoblast apoptosis in osteoporosis by attenuating the maturation of miR-1224-5p. Journal of orthopaedic surgery and research 5 38183099
2024 E3 ubiquitin ligase HECW2: a promising target for tumour therapy. Cancer cell international 5 39529070
2016 Decreased expression of NEDL2 in Hirschsprung's disease. Journal of pediatric surgery 5 27430863
2022 First splicing variant in HECW2 with an autosomal recessive pattern of inheritance and associated with NDHSAL. Human mutation 3 35753050
2025 HECW2 knockdown suppresses the development of infantile hemangioma by inhibiting ALKBH5/LDHA axis-mediated glycolysis. American journal of cancer research 2 40520873
2022 A De Novo HECW2 Variant in a Patient with Acetazolamide-Responsive Episodic Ataxia. Cerebellum (London, England) 2 35987951
2025 Validation of GDAP1 and HECW2 as Epigenetic Markers of Alcohol Use Disorder in Blood and Brain. International journal of molecular sciences 1 41303328
2025 A Novel HECW2 Variant (c.4354G>A; p. Gly1452Ser) in a Chinese Patient with Developmental Delay, Neurodevelopmental Delay, and Hypotonia. Molecular syndromology 0 40469502
2025 HECW2 Gene Mutation: A Rare Cause of West Syndrome: A Case Report. Neurology India 0 40705299
2025 Autosomal recessive frameshift variant broadens HECW2-related disease spectrum. European journal of medical genetics 0 40812465
2025 Rapid Electroclinical Evolution in HECW2-Related Developmental and Epileptic Encephalopathy: Report of a Likely Splicing Variant With Familial Transmission. International journal of developmental neuroscience : the official journal of the International Society for Developmental Neuroscience 0 40879666

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