Affinage

CHFR

E3 ubiquitin-protein ligase CHFR · UniProt Q96EP1

Length
664 aa
Mass
73.4 kDa
Annotated
2026-06-09
100 papers in source corpus 35 papers cited in narrative 35 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 8/8 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

CHFR is a RING-domain E3 ubiquitin ligase and tumor suppressor that enforces a mitotic stress (antephase/prophase) checkpoint, delaying chromosome condensation in response to microtubule poisons (PMID:10935642). Its checkpoint activity is genetically separable from its catalytic activity: the FHA domain mediates growth suppression and checkpoint function while the RING and cysteine-rich regions are required for ubiquitin ligase activity (PMID:18335050). The principal effector of the checkpoint is Polo-like kinase 1 (Plk1), which CHFR ubiquitinates to prolong inhibitory Cdc2 tyrosine-15 phosphorylation by delaying Cdc25C activation and Wee1 inactivation, thereby excluding Cyclin B1 from the nucleus and blocking mitotic entry (PMID:11807090, PMID:15674323, PMID:21768102), a checkpoint that operates downstream of the p38 stress kinases (PMID:15302856). CHFR builds distinct chain types with distinct E2s — Lys48-linked chains via Ubc4/Ubc5 that target substrates for proteasomal degradation, and non-degradative Lys63-linked chains via Ubc13-Mms2 for stress signaling (PMID:11912157, PMID:14562038, PMID:18202552). Beyond Plk1, CHFR ubiquitinates a broad substrate set to maintain chromosomal stability and regulate transcription, migration, and apoptosis, including Aurora A (PMID:15793587), HDAC1 (PMID:19182791), Kif22 (PMID:19321445), PARP1 (PMID:23268447, PMID:22337872), HLTF (PMID:20388495), SIRT1 (PMID:27883020), ZEB1 (PMID:34462429), SWI/SNF-BAF components (PMID:22285184), RNF126 (PMID:34388456), and VE-cadherin in endothelial barrier control (PMID:37852964). CHFR is recruited to DNA double-strand breaks through poly(ADP-ribose) binding by its cysteine-rich PAR-binding zinc finger (PBZ), whose structure and PAR-recognition mode are defined, where it directs the first wave of damage-site ubiquitination and cooperates with RNF8 to control histone H4K16 acetylation and ATM activation (PMID:20880844, PMID:21706008, PMID:23268447). CHFR activity is set by counter-regulators: USP7/HAUSP deubiquitination stabilizes it (PMID:17442268), STIL promotes its auto-ubiquitination and turnover (PMID:21245198), and PKB/Akt phosphorylation inhibits its checkpoint function (PMID:14638868). Loss of CHFR causes chromosomal instability and cancer predisposition in knockout mice (PMID:15793587, PMID:21706008).

Mechanistic history

Synthesis pass · year-by-year structured walk · 23 steps
  1. 2000 High

    Established that CHFR exists and defines a previously unrecognized mitotic stress checkpoint controlling the timing of chromosome condensation.

    Evidence Ectopic wild-type vs. mutant CHFR expression and cell-cycle analysis after microtubule poisons in cancer cell lines

    PMID:10935642

    Open questions at the time
    • Molecular activity of CHFR unknown at this stage
    • Direct substrates unidentified
  2. 2002 High

    Defined CHFR as a RING-domain E3 ubiquitin ligase and identified Plk1 as the checkpoint substrate, explaining how the checkpoint delays Cdc2 activation.

    Evidence In vitro ubiquitination, RING mutagenesis, Xenopus extracts, and proteasome inhibition; independent confirmation of RING requirement and cell-cycle-regulated expression

    PMID:11807090 PMID:11912157

    Open questions at the time
    • Chain linkage type not yet resolved
    • Whether Plk1 degradation is the sole mechanism unclear
  3. 2002 High

    Determined the FHA domain structure, indicating a phosphopeptide-recognition module consistent with FHA family proteins.

    Evidence X-ray crystallography of FHA domain native and tungstate-bound forms

    PMID:12121644

    Open questions at the time
    • Physiological phosphoprotein ligand of the FHA domain not identified
    • Crystallized as segment-swapped dimer of uncertain in vivo relevance
  4. 2003 High

    Showed CHFR can build non-degradative Lys63-linked chains via Ubc13-Mms2, distinguishing a signaling function from proteasomal targeting.

    Evidence In vitro ubiquitination with Ubc13-Mms2 and chain linkage analysis; mitotic mobility-shift analysis

    PMID:14562038

    Open questions at the time
    • In vivo substrates of K63 chains not defined
    • Reconciliation with degradative K48 activity left open
  5. 2004 High

    Placed CHFR downstream of p38 stress kinases and confirmed the antephase checkpoint requires ubiquitylation but not the proteasome.

    Evidence p38 inhibition, checkpoint assays in Chfr-deficient cells, proteasome inhibitor experiments

    PMID:15302856

    Open questions at the time
    • Direct link between p38 and CHFR activation not biochemically defined
  6. 2004 High

    Linked CHFR nuclear dynamics and checkpoint function to PML bodies.

    Evidence Live-cell FRET, fluorescence recovery, PML-knockout cells, checkpoint assays

    PMID:15467728

    Open questions at the time
    • Mechanism by which PML bodies regulate CHFR activity unresolved
  7. 2005 High

    Identified Aurora A as a substrate and established CHFR as a chromosomal-stability tumor suppressor in vivo.

    Evidence Co-IP, in vitro ubiquitination, Chfr-knockout mice, MEF chromosome analysis

    PMID:15793587

    Open questions at the time
    • Relative contribution of Aurora A vs. Plk1 to tumor suppression unclear
  8. 2005 High

    Defined the downstream output of the checkpoint as nuclear exclusion of Cyclin B1, preventing chromosome condensation.

    Evidence Wild-type CHFR cell lines, cell-cycle marker immunofluorescence, Cyclin B1 NES-mutant override experiment

    PMID:15674323

    Open questions at the time
    • Mechanism connecting Plk1 ubiquitination to Cyclin B1 localization not fully traced
  9. 2008 Medium

    Separated CHFR's checkpoint function (FHA domain) from its catalytic activity (RING + cysteine-rich domains).

    Evidence Domain deletion mutagenesis with proliferation, checkpoint, and ubiquitination assays

    PMID:18335050

    Open questions at the time
    • How FHA-dependent checkpoint function operates without requiring ligase activity not explained
    • Single lab
  10. 2008 Medium

    Localized endogenous CHFR to the cytoplasm and mitotic spindle and proposed microtubule sensing through TCTP/tubulin interactions.

    Evidence Two-hybrid screen, endogenous Co-IP, immunofluorescence with three antibodies, nocodazole depolymerization

    PMID:18504434

    Open questions at the time
    • Causal role of TCTP/tubulin binding in checkpoint activation not demonstrated
    • Ectopic overexpression shifts localization, complicating interpretation
  11. 2008 Medium

    Extended CHFR's spindle role through alpha-tubulin and MAD2 regulation affecting spindle assembly checkpoint integrity.

    Evidence siRNA knockdown, Co-IP, ubiquitination assays, mitotic spindle microscopy, chromosome counting

    PMID:18592005

    Open questions at the time
    • Direct vs. indirect effects on MAD2/BUBR1 localization not distinguished
    • Single lab
  12. 2009 Medium

    Expanded the substrate repertoire to transcriptional and motility regulators (HDAC1, Kif22, HLTF), linking CHFR to gene expression, invasion, and chromosome stability.

    Evidence AP-MS, Co-IP, in vitro ubiquitination, invasion/migration assays, xenograft model

    PMID:19182791 PMID:19321445 PMID:20388495

    Open questions at the time
    • Physiological conditions selecting each substrate undefined
    • Some readouts rely on overexpression
  13. 2009 Medium

    Identified upstream regulation by PKB/Akt phosphorylation that inhibits checkpoint function and a KKK NLS required for nuclear checkpoint activity.

    Evidence In vitro kinase assay, phospho-mutant rescue, NLS mutagenesis, localization and checkpoint assays

    PMID:14638868 PMID:19326084

    Open questions at the time
    • Phosphosite mapping limited
    • Single-lab findings
  14. 2010 High

    Defined the PBZ zinc finger structure and its PAR-recognition mode, establishing the molecular basis for PAR-dependent recruitment and antephase checkpoint function.

    Evidence 1.9 Å crystal and co-crystal structures with PAR analogs, mutagenesis, SPR binding validation

    PMID:20880844

    Open questions at the time
    • In vivo PAR sources driving CHFR recruitment in different contexts not fully mapped
  15. 2011 High

    Established CHFR's role in the early DNA damage response: PAR-dependent recruitment to DSBs, PARP1 ubiquitination, and RNF8-cooperative control of H4K16 acetylation and ATM activation.

    Evidence Laser-induced DSB recruitment, PAR binding, in vivo ubiquitination, RNF8/Chfr double-knockout mice, histone modification and ATM assays

    PMID:21706008 PMID:23268447

    Open questions at the time
    • Order of events between CHFR and RNF8 at chromatin not fully resolved
  16. 2011 High

    Clarified the role of CHFR auto-ubiquitylation in normal cell-cycle timing, showing auto-ubiquitylation-driven turnover permits Plk1 accumulation for mitotic entry, with Plk1 the dominant target over Aurora A.

    Evidence Auto-ubiquitylation site mutants, cell-cycle analysis, Plk1/Aurora A measurement, Cyclin B1/Cdk1 kinase assay, Plk1 rescue

    PMID:21768102

    Open questions at the time
    • Trigger controlling timing of auto-ubiquitylation not defined
  17. 2011 Medium

    Identified STIL as the first negative regulator promoting CHFR auto-ubiquitination and turnover.

    Evidence Stil-knockout mice/MEFs, siRNA, stability assays, rescue by Chfr KD or Plk1 OE

    PMID:21245198

    Open questions at the time
    • Whether STIL acts directly on CHFR not established
    • Single lab
  18. 2012 High

    Showed PARP1 auto-PARylation under mitotic stress enhances CHFR-PARP1 interaction and PARP1 degradation, coupling PAR signaling to checkpoint arrest.

    Evidence AP-MS, Co-IP, in vitro ubiquitination, Chfr-KO mouse tissue, cell-cycle assays

    PMID:22337872

    Open questions at the time
    • Quantitative contribution of PARP1 degradation to arrest vs. other substrates unclear
  19. 2012 Medium

    Connected CHFR to additional checkpoint and metabolic signaling via TOPK/PTEN/Akt and BAF complex stabilization by SRG3/mBAF155.

    Evidence Co-IP, in vitro ubiquitination, kinase assays, protein stability assays

    PMID:22285184 PMID:24012691

    Open questions at the time
    • Pathway placements rest on epistasis in single labs
    • Physiological substrate selection conditions undefined
  20. 2015 Medium

    Demonstrated that PBZ-mediated CHFR-PARP1 interaction stabilizes CHFR and is a druggable node, with a PBZ-targeting compound synergizing with docetaxel.

    Evidence shRNA, PBZ peptide overexpression, docking screen, protein-interaction and viability assays, xenograft model

    PMID:26356822

    Open questions at the time
    • Compound specificity and off-target effects not fully characterized
    • Single lab
  21. 2016 Medium

    Identified SIRT1 as an oxidative-stress substrate, linking CHFR to JNK1-dependent p53 acetylation and apoptosis.

    Evidence Co-IP, in vivo ubiquitination, stability assays, JNK inhibition, p53 acetylation assay

    PMID:27883020

    Open questions at the time
    • Direct ubiquitination not reconstituted in vitro
    • Single lab
  22. 2021 Medium

    Extended CHFR substrate range to ZEB1 (chemo-sensitization in TNBC) and RNF126 (DNA damage checkpoint modulation), the latter via PAR-dependent recruitment.

    Evidence AP-MS, Co-IP, in vivo/in vitro ubiquitination, PAR-binding assays, viability assays

    PMID:34388456 PMID:34462429

    Open questions at the time
    • Context-dependence of these substrate relationships not defined
    • Single-lab studies
  23. 2023 High

    Established a non-mitotic physiological role for CHFR in endothelial barrier integrity through VE-cadherin degradation, downstream of FoxO1.

    Evidence Endothelial-specific conditional Chfr knockout mice, immunofluorescence, VE-cadherin ubiquitination assay, LPS challenge, permeability assays

    PMID:37852964

    Open questions at the time
    • Whether VE-cadherin is a direct ubiquitination substrate in vitro not shown
    • Link to mitotic functions unclear

Open questions

Synthesis pass · forward-looking unresolved questions
  • How CHFR integrates upstream stress signals (p38, JNK, Akt, PAR) to select among its many substrates and switch between K48 and K63 chain outputs in specific contexts remains unresolved.
  • No unified model linking signal input to substrate/chain-type choice
  • FHA domain physiological phospholigand unidentified
  • Mechanism coupling cytoplasmic microtubule sensing to nuclear checkpoint output incomplete

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0140096 catalytic activity, acting on a protein 8 GO:0016874 ligase activity 7 GO:0003723 RNA binding 3
Localization
GO:0000228 nuclear chromosome 2 GO:0005634 nucleus 2 GO:0005856 cytoskeleton 2 GO:0005829 cytosol 1
Pathway
R-HSA-1640170 Cell Cycle 4 R-HSA-392499 Metabolism of proteins 3 R-HSA-73894 DNA Repair 3 R-HSA-4839726 Chromatin organization 1
Partners
AURKAHDAC1MAD2L1PARP1PLK1RNF8TPT1USP7

Evidence

Reading pass · 35 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2000 CHFR defines a mitotic stress checkpoint that delays entry into metaphase (prophase checkpoint). Cells expressing wild-type CHFR delay chromosome condensation in response to microtubule poisons (nocodazole, taxol); cells lacking CHFR expression enter metaphase without delay. Ectopic expression of wild-type CHFR restored the cell cycle delay and increased cell survival under mitotic stress. Ectopic expression of wild-type vs. mutant CHFR in human cancer cell lines; cell cycle analysis after microtubule poison treatment Nature High 10935642
2002 CHFR is an E3 ubiquitin ligase; its RING finger domain is required for ubiquitin ligase activity. Chfr auto-ubiquitinates in vitro and in vivo, and mutations of conserved RING-finger residues abolish ligase activity. Chfr delays mitotic entry by negatively regulating Cdc2 kinase activation at the G2-M transition by prolonging phosphorylation of Cdc2 tyrosine 15. The direct target of the Chfr pathway is Polo-like kinase 1 (Plk1): Chfr ubiquitinates Plk1, delaying activation of Cdc25C and inactivation of Wee1, thereby delaying Cdc2 activation. The Chfr pathway requires ubiquitin-dependent protein degradation (inactivating RING-domain mutations, poly-ubiquitination interference, or proteasome inhibition all abolish the delay). In vitro ubiquitination assays; in vivo overexpression in HEK293T; Xenopus cell-free extracts; RING-domain mutagenesis; proteasome inhibitor experiments The Journal of cell biology High 11807090
2002 CHFR RING finger domain is required for ubiquitin ligase activity (auto-ubiquitination in vitro with E1, Ubc4/Ubc5, and ubiquitin). RING domain deletion mutant is defective in inhibiting chromosome condensation after taxol treatment. CHFR expression is cell cycle regulated and dependent on its ubiquitin ligase activity. In vitro ubiquitination assay with purified components; stable cell lines expressing wild-type or RING-deleted CHFR; taxol treatment cell cycle assay Cancer research High 11912157
2002 Crystal structure of the CHFR FHA domain (native and in complex with tungstate, a phosphate analog) revealed a beta-sandwich fold similar to Rad53 FHA domains but crystallizing as a segment-swapped dimer. The ability to recognize tungstate suggests the CHFR FHA domain shares the ability of other FHA domains to bind phosphoproteins, though sequence/structure differences indicate distinct binding properties across FHA domain families. X-ray crystallography (structure determination of FHA domain native and tungstate-bound forms) Structure (London, England : 1993) High 12121644
2003 CHFR catalyzes the formation of non-canonical Lys63-linked polyubiquitin chains using Ubc13-Mms2 as the ubiquitin-conjugating enzyme. Ubc13-Mms2-dependent Lys63 chains are not associated with proteasomal targeting but with stress signaling. In mitosis, Chfr undergoes a phosphorylation-dependent electrophoretic mobility shift with no change in overall protein levels (contrary to a prior report suggesting Chfr undergoes proteasome-dependent degradation in mitosis). In vitro ubiquitination assay with Ubc13-Mms2; polyubiquitin chain linkage analysis; cell cycle protein level analysis Oncogene High 14562038
2004 In mammalian cells, the antephase checkpoint (blocking mitotic entry in response to microtubule poisons) is primarily mediated by the p38 stress kinases and requires the Chfr protein. The checkpoint requires ubiquitylation but not proteasome activity, consistent with Chfr forming Lys63-linked (non-degradative) ubiquitin chains. Genetic/pharmacological inhibition of p38 kinases; checkpoint assays in Chfr-deficient vs. Chfr-expressing cell lines; proteasome inhibitor experiments The Journal of cell biology High 15302856
2004 PML bodies control the nuclear dynamics and function of CHFR. CHFR is a dynamic component of PML bodies; FRET analysis identified a fraction of CHFR that interacts with PML in living cells. A trans-dominant CHFR mutant that inhibits checkpoint function also prevents colocalization/interaction with PML. In PML−/− cells, CHFR distribution and mobility are perturbed, accompanied by aberrations in mitotic entry and response to spindle depolymerization. Live-cell FRET; fluorescence recovery analysis; PML-knockout cells; checkpoint assays Nature structural & molecular biology High 15467728
2005 CHFR physically interacts with Aurora A and ubiquitinates Aurora A both in vitro and in vivo, controlling Aurora A expression levels. Chfr-deficient mice are cancer-prone with chromosomal instability in embryonic fibroblasts, establishing Chfr as a tumor suppressor that maintains chromosomal stability partly through control of Aurora A. Co-immunoprecipitation; in vitro ubiquitination assay; Chfr knockout mouse model; MEF chromosome analysis Nature genetics High 15793587
2005 CHFR delays chromosome condensation in prophase by excluding Cyclin B1 from the nucleus. In CHFR-expressing cells arrested by microtubule poisons, Cyclin A/Cdc2 is activated but Aurora-A, Aurora-B, and Cyclin B1/Cdc2 are inactive; Cyclin B1 is excluded from the nucleus. Ectopic expression of Cyclin B1 with a mutant nuclear export sequence induces chromosome condensation and overrides the CHFR checkpoint. Stable cell lines expressing wild-type CHFR; immunofluorescence and cell cycle marker analysis after microtubule poison treatment; ectopic Cyclin B1 nuclear export mutant expression Oncogene High 15674323
2007 USP7/HAUSP is a deubiquitinase that interacts with Chfr and removes ubiquitin moieties from auto-ubiquitinated Chfr both in vivo and in vitro, resulting in Chfr accumulation. USP7-mediated deubiquitination of Chfr leads to its stabilization and may be a key regulatory step for Chfr activation. Immunoaffinity purification and mass spectrometry; co-immunoprecipitation; in vitro deubiquitination assay Biochemical and biophysical research communications Medium 17442268
2008 Endogenous CHFR localizes to the cytoplasm and to the mitotic spindle during mitosis. CHFR interacts with TCTP and beta-tubulin (confirmed by biochemical analysis of endogenous proteins). The Chfr-TCTP interaction is stable throughout the cell cycle but is diminished by complete depolymerization of microtubules, suggesting Chfr may sense microtubule disruption through this interaction to activate the prophase checkpoint. High-level ectopic Chfr overexpression shifts localization to the nucleus/PML bodies and causes protein degradation in an autoubiquitination/proteasome-dependent manner. Two-hybrid screen; co-immunoprecipitation of endogenous proteins; immunofluorescence with three different anti-Chfr antibodies; nocodazole microtubule depolymerization assay Oncogene Medium 18504434
2008 The FHA domain of CHFR is critical for its anti-proliferative and G2/M checkpoint functions; the RING finger and cysteine-rich domains are required for E3 ligase activity but not for growth suppression. Domain deletion analysis established that checkpoint function (FHA domain) and ubiquitin ligase activity (RING + Cys domains) are separable activities. Domain deletion mutagenesis; cell proliferation assays; cell cycle checkpoint assays; in vitro/in vivo ubiquitination assays PloS one Medium 18335050
2008 CHFR interacts with alpha-tubulin and can regulate its ubiquitination in response to nocodazole and control the amount of acetylated alpha-tubulin (a spindle component). CHFR also interacts with the spindle checkpoint protein MAD2; decreased CHFR expression results in mislocalization of MAD2 and BUBR1 during mitosis and impaired MAD2/CDC20 complex formation, leading to multipolar spindles and genomic instability. siRNA knockdown of CHFR in MCF10A; co-immunoprecipitation (CHFR with Aurora A, alpha-tubulin, MAD2); ubiquitination assays; fluorescence microscopy of mitotic spindle; chromosome counting Neoplasia (New York, N.Y.) Medium 18592005
2009 CHFR binds and ubiquitinates HDAC1 (histone deacetylase 1) both in vitro and in vivo, targeting it for degradation. Ectopic expression of CHFR in cancer cells leads to downregulation of HDAC1, upregulation of p21(CIP1/WAF1), KAI1, and E-cadherin, G1 arrest, and reduced invasiveness. Identified via affinity purification/mass spectrometry. Affinity purification combined with mass spectrometry; co-immunoprecipitation; in vitro ubiquitination assay; invasion assays; xenograft model Nature cell biology High 19182791
2009 CHFR interacts with Kif22 (a chromokinesin that binds DNA and microtubules) and ubiquitinates Kif22 as a novel substrate, leading to Kif22 downregulation. Chfr-mediated Kif22 down-regulation is critical for the maintenance of chromosome stability. Co-immunoprecipitation; in vitro ubiquitination assay; siRNA knockdown; chromosome stability assays The Journal of biological chemistry Medium 19321445
2009 Activated PKB/Akt phosphorylates CHFR in vitro and in vivo. Expression of a CHFR mutant that cannot be phosphorylated by PKB results in reduced Plk1 levels and inhibition of mitotic entry. Thus, PKB facilitates mitotic entry after DNA damage by phosphorylating and inhibiting CHFR checkpoint function. In vitro kinase assay (PKB phosphorylating CHFR); phospho-mutant CHFR expression; Plk1 level analysis; mitotic entry assay Molecular cancer research : MCR Medium 14638868
2009 Nuclear localization of CHFR is mediated by a nuclear localization signal (NLS); a lysine-rich stretch (KKK) at amino acids 257-259 is essential for nuclear localization. Nuclear localization is essential for CHFR checkpoint function but not for its stability. NLS mutagenesis (KKK→AAA); immunofluorescence localization; checkpoint functional assays Molecules and cells Medium 19326084
2010 Crystal structure of the cysteine-rich region of human CHFR (residues 425–664) at 1.9 Å resolution revealed a multizinc binding domain with elaborate topology containing the PAR-binding zinc finger (PBZ). Co-crystal structures with PAR-like ligands (ADP-ribose, AMP, diadenosine pyrophosphate) defined the PBZ-PAR interaction: the PBZ recognizes two adenine-containing subunits of PAR and the connecting phosphate backbone. Site-specific mutagenesis and surface plasmon resonance validated this interaction. The PBZ-PAR interaction is required for antephase checkpoint function. X-ray crystallography (1.9 Å); co-crystal structures with PAR analogs; site-specific mutagenesis; surface plasmon resonance The Journal of biological chemistry High 20880844
2010 CHFR functions as a ubiquitin ligase for HLTF (helicase-like transcription factor), binding to and ubiquitinating HLTF to target it for proteasomal degradation. CHFR-mediated HLTF downregulation reduces PAI-1 expression, inhibiting cell migration. Co-immunoprecipitation; in vitro ubiquitination assay; cell migration assay Biochemical and biophysical research communications Medium 20388495
2011 STIL promotes Chfr auto-ubiquitination and reduces its protein stability. In cells lacking STIL (MEFs, E9.5 embryos), Chfr protein levels are elevated and Plk1 (a Chfr substrate) is reduced. Knockdown of Chfr or overexpression of Plk1 reverses the abnormal mitotic phenotypes of STIL-deficient fibroblasts. STIL is thus identified as the first negative regulator of the Chfr mitotic checkpoint. Genetic KO (Stil-/- mice); siRNA knockdown; protein stability assays; rescue by Chfr KD or Plk1 OE; flow cytometry Journal of cell science Medium 21245198
2011 RNF8 and CHFR synergistically regulate histone ubiquitination to control histone H4 Lys16 acetylation through MRG15-dependent acetyltransferase complexes. This chromatin remodeling function is required for ATM activation after DNA damage. Double-knockout (RNF8/Chfr DKO) cells show suppressed DNA damage-induced ATM kinase activation, hypersensitivity to ionizing radiation, and DKO mice develop thymic lymphomas with clonal chromosome translocations. Double-knockout mouse model; chromatin fractionation; histone ubiquitination and acetylation assays; ATM activation assays; radiation sensitivity assays Nature structural & molecular biology High 21706008
2011 Auto-ubiquitylation of Chfr at G2 phase is required for Plk1 accumulation and mitotic entry. Chfr auto-ubiquitylation mutants (Chfr-K2A) do not undergo cell cycle-dependent degradation; elevated Chfr-K2A levels at G2/M reduce Plk1 levels and delay mitotic entry by suppressing cyclin B1/Cdk1 kinase activity. Overexpression of Plk1 in Chfr-K2A cells restores kinase activity and promotes mitotic entry. Aurora A levels are not affected, identifying Plk1 as the major Chfr target in controlling mitotic entry timing. Site-directed mutagenesis of Chfr auto-ubiquitylation sites; cell cycle analysis; Plk1 and Aurora A level measurement; cyclin B1/Cdk1 kinase assay; Plk1 overexpression rescue The Journal of biological chemistry High 21768102
2011 CHFR is recruited to DNA double-strand breaks (DSBs) via poly(ADP-ribose) (PAR). At DSBs, CHFR regulates the first wave of protein ubiquitination. CHFR ubiquitinates PARP1 and regulates chromatin-associated PARP1 in vivo, establishing CHFR as an important E3 ligase in the early DNA damage response that mediates crosstalk between ubiquitination and poly-ADP-ribosylation. Recruitment assay to laser-induced DSBs; PAR-binding experiments; in vivo ubiquitination assay; chromatin fractionation Nucleic acids research Medium 23268447
2011 CHFR binds MAD2 via its cysteine-rich domain; deletion of this domain abolishes the CHFR/MAD2 interaction. The cysteine-rich domain is required for proper MAD2 localization and for promoting the MAD2/CDC20 interaction, thus enabling spindle assembly checkpoint function. Domain deletion mutagenesis; co-immunoprecipitation; MAD2 localization by immunofluorescence; MAD2/CDC20 interaction assay Biochemical and biophysical research communications Medium 21575600
2012 CHFR interacts with PARP-1; mitotic stress induces PARP-1 auto-PARylation, which enhances the CHFR-PARP-1 interaction and increases polyubiquitination/degradation of PARP-1. Decreased PARP-1 protein levels promote cell cycle arrest at prophase. In CHFR-silenced cells, PARP-1 is not degraded after mitotic stress and cells progress into mitosis. Chfr knockout mice and CHFR-silenced primary gastric cancer tissues have higher PARP-1 protein levels. Affinity purification/mass spectrometry; co-immunoprecipitation; in vitro ubiquitination; Chfr KO mouse tissue analysis; cell cycle assays The Journal of biological chemistry High 22337872
2012 SRG3/mBAF155 blocks CHFR-mediated ubiquitination and degradation of core SWI/SNF-BAF complex components (BRG1, SNF5, BAF60a). CHFR interacts with BRG1, SNF5, and BAF60a and ubiquitinates them for proteasomal degradation; SRG3/mBAF155 stabilizes these components by blocking their interaction with CHFR. Co-immunoprecipitation; in vitro ubiquitination assay; protein stability assays Biochemical and biophysical research communications Medium 22285184
2013 CHFR ubiquitinates and regulates TOPK/PBK (a serine/threonine kinase) levels as part of its checkpoint function. TOPK in turn phosphorylates and inactivates PTEN, which activates Akt, leading to proper G2/M progression. TOPK and PTEN are identified as new players in CHFR-mediated mitotic checkpoint signaling. Co-immunoprecipitation; in vitro ubiquitination assay; kinase assay (TOPK phosphorylation of PTEN); CHFR knockdown/overexpression Cellular signalling Medium 24012691
2015 The CHFR PBZ domain mediates a protein interaction with poly-ADP-ribosylated PARP1, leading to stabilization of CHFR. Disruption of the CHFR-PARP1 interaction (via PARP1 knockdown or PBZ peptide overexpression) causes loss of CHFR protein expression. A small-molecule inhibitor ('A3') targeting the CHFR PBZ domain disrupts the CHFR-PARP1 interaction and inhibits mitotic checkpoint function, showing therapeutic synergy with docetaxel in vitro and in xenograft models. shRNA knockdown; PBZ domain peptide overexpression; computational docking screen; in vitro protein-protein interaction assay; cell viability and colony formation assays; xenograft mouse model Oncotarget Medium 26356822
2016 CHFR functions as an E3 ubiquitin ligase for SIRT1, responsible for its proteasomal degradation under oxidative stress. CHFR interacts with SIRT1 and ubiquitinates it. CHFR-mediated SIRT1 degradation is enhanced when SIRT1 is phosphorylated by JNK1 upon oxidative stress, leading to increased p53 acetylation and apoptosis. JNK inhibition prevents SIRT1 phosphorylation and CHFR-mediated degradation. Co-immunoprecipitation; in vivo ubiquitination assay; protein stability assay; JNK inhibitor experiments; p53 acetylation assay Scientific reports Medium 27883020
2021 CHFR associates with and ubiquitinates ZEB1 in a ubiquitination-dependent manner, leading to decreased ZEB1 expression in TNBC. CHFR represses FASN expression through ZEB1, causing cell death under chemotherapy. HDAC inhibitor TSA increases CHFR expression independent of histone acetylation, destabilizing ZEB1 and sensitizing TNBC cells to chemotherapy. Affinity purification/mass spectrometry; co-immunoprecipitation; in vivo ubiquitination assay; CHFR overexpression/knockdown; cell viability assays Cell death & disease Medium 34462429
2021 PARP1 interacts with and poly(ADP-ribosyl)ates RNF126, which then recruits CHFR (via its PAR-binding PBZ domain) to ubiquitinate and degrade RNF126. RNF126 is required for ATR-Chk1 signaling activation by IR or PARP inhibitors; CHFR-mediated RNF126 degradation thus modulates DNA damage checkpoint signaling. Co-immunoprecipitation; in vitro and in vivo ubiquitination assay; PAR-binding assays; siRNA knockdown; cell viability assays Biochemical and biophysical research communications Medium 34388456
2023 CHFR is required for ubiquitylation-dependent degradation of VE-cadherin in endothelial cells. Endothelial-restricted Chfr knockout (ChfrΔEC) mice show augmented VE-cadherin expression at adherens junctions and abrogated LPS-induced VE-cadherin degradation. LPS-induced lung endothelial barrier breakdown, inflammatory neutrophil extravasation, and mortality are suppressed in ChfrΔEC mice. FoxO1 transcription factor is identified as an upstream regulator of CHFR expression. Endothelial-specific conditional Chfr knockout mice; immunofluorescence; VE-cadherin ubiquitination assay; LPS challenge model; endothelial permeability assay Nature communications High 37852964
2007 In S. cerevisiae Chfr homologs (Chf1/Chf2), Ubc4 is required for G1 cell cycle delay and Chf protein turnover (Lys48-type modification), while Ubc13/Mms2 is required for G2 delay and does not contribute to protein turnover (Lys63-type modification). Site-specific mutagenesis identified the Lys residues modified by each E2. This dual E2 specificity accounts for different cell cycle functions. Yeast genetic assays; in vitro reconstitution with purified components; tandem mass spectrometry linkage analysis; site-directed mutagenesis Cell cycle (Georgetown, Tex.) High 18202552
2004 S. cerevisiae Dma1 and Dma2 (budding yeast Chfr homologs) are involved in proper spindle positioning, likely by regulating septin ring deposition at the bud neck. DMA2 overexpression causes defects in septin ring disassembly and cytokinesis that can be rescued by eliminating the spindle position checkpoint protein Bub2 or overproducing Tem1. dma1Δdma2Δ cells fail to activate the spindle position checkpoint in response to dynein loss. Genetic deletion analysis; DMA2 overexpression; epistasis with Bub2/Tem1; spindle position checkpoint assays Molecular biology of the cell Medium 15146058
2012 CHFR interacts with LRP1 in human vascular smooth muscle cells. Aggregated LDL (agLDL) downregulates CHFR expression and reduces the CHFR-LRP1 physical interaction, leading to reduced LRP1 ubiquitination and prolonged LRP1 half-life. CHFR silencing increases VSMC LRP1 expression specifically (not classical LDLR), indicating CHFR targets LRP1 for ubiquitin-mediated degradation. Co-immunoprecipitation; pulse-chase analysis; cycloheximide chase; CHFR siRNA silencing; 2D proteomics Arteriosclerosis, thrombosis, and vascular biology Medium 23241409

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2000 Chfr defines a mitotic stress checkpoint that delays entry into metaphase. Nature 315 10935642
2003 Epigenetic inactivation of CHFR in human tumors. Proceedings of the National Academy of Sciences of the United States of America 174 12810945
2002 The checkpoint protein Chfr is a ligase that ubiquitinates Plk1 and inhibits Cdc2 at the G2 to M transition. The Journal of cell biology 172 11807090
2005 Chfr is required for tumor suppression and Aurora A regulation. Nature genetics 169 15793587
2003 Epigenetic inactivation of CHFR and sensitivity to microtubule inhibitors in gastric cancer. Cancer research 145 14695171
2002 Aberrant hypermethylation of the CHFR prophase checkpoint gene in human lung cancers. Oncogene 111 11948416
2004 Chfr acts with the p38 stress kinases to block entry to mitosis in mammalian cells. The Journal of cell biology 93 15302856
2003 Frequent hypermethylation of the 5' CpG island of the mitotic stress checkpoint gene Chfr in colorectal and non-small cell lung cancer. Carcinogenesis 90 12538348
2011 Chfr and RNF8 synergistically regulate ATM activation. Nature structural & molecular biology 87 21706008
2002 Chfr expression is downregulated by CpG island hypermethylation in esophageal cancer. Carcinogenesis 85 12376479
2012 CHFR protein regulates mitotic checkpoint by targeting PARP-1 protein for ubiquitination and degradation. The Journal of biological chemistry 83 22337872
2002 Chfr regulates a mitotic stress pathway through its RING-finger domain with ubiquitin ligase activity. Cancer research 80 11912157
2003 The Chfr mitotic checkpoint protein functions with Ubc13-Mms2 to form Lys63-linked polyubiquitin chains. Oncogene 79 14562038
2009 Chfr is linked to tumour metastasis through the downregulation of HDAC1. Nature cell biology 75 19182791
2005 CHFR promoter hypermethylation in colon cancer correlates with the microsatellite instability phenotype. Carcinogenesis 73 15760919
2012 CHFR is important for the first wave of ubiquitination at DNA damage sites. Nucleic acids research 67 23268447
2014 Predictive value of CHFR and MLH1 methylation in human gastric cancer. Gastric cancer : official journal of the International Gastric Cancer Association and the Japanese Gastric Cancer Association 66 24748501
2010 Structural basis of poly(ADP-ribose) recognition by the multizinc binding domain of checkpoint with forkhead-associated and RING Domains (CHFR). The Journal of biological chemistry 58 20880844
2005 The CHFR mitotic checkpoint protein delays cell cycle progression by excluding Cyclin B1 from the nucleus. Oncogene 53 15674323
2008 Chfr interacts and colocalizes with TCTP to the mitotic spindle. Oncogene 51 18504434
2006 The significance of aberrant CHFR methylation for clinical response to microtubule inhibitors in gastric cancer. Journal of gastroenterology 46 16568372
2004 Functional characterization of Dma1 and Dma2, the budding yeast homologues of Schizosaccharomyces pombe Dma1 and human Chfr. Molecular biology of the cell 46 15146058
2020 Circ_CHFR expedites cell growth, migration and inflammation in ox-LDL-treated human vascular smooth muscle cells via the miR-214-3p/Wnt3/β-catenin pathway. European review for medical and pharmacological sciences 44 32271446
2007 Deubiquitination of Chfr, a checkpoint protein, by USP7/HAUSP regulates its stability and activity. Biochemical and biophysical research communications 44 17442268
2005 Epigenetic inactivation of CHFR in nasopharyngeal carcinoma through promoter methylation. Molecular carcinogenesis 43 15937956
2002 Crystal structure of the FHA domain of the Chfr mitotic checkpoint protein and its complex with tungstate. Structure (London, England : 1993) 43 12121644
2003 Inactivating mutations targeting the chfr mitotic checkpoint gene in human lung cancer. Cancer research 42 14612512
2011 The Stil protein regulates centrosome integrity and mitosis through suppression of Chfr. Journal of cell science 41 21245198
2003 Promotion of mitosis by activated protein kinase B after DNA damage involves polo-like kinase 1 and checkpoint protein CHFR. Molecular cancer research : MCR 41 14638868
2007 Altered expression of the early mitotic checkpoint protein, CHFR, in breast cancers: implications for tumor suppression. Cancer research 40 17596595
2005 Small interfering RNA-induced CHFR silencing sensitizes oral squamous cell cancer cells to microtubule inhibitors. Cancer biology & therapy 39 16123600
2013 TOPK and PTEN participate in CHFR mediated mitotic checkpoint. Cellular signalling 37 24012691
2008 CHFR: A Novel Mitotic Checkpoint Protein and Regulator of Tumorigenesis. Translational oncology 36 18633460
2004 Promoter hypermethylation of the Chfr gene in neoplastic and non-neoplastic gastric epithelia. British journal of cancer 36 15138487
2014 Emerging evidence for CHFR as a cancer biomarker: from tumor biology to precision medicine. Cancer metastasis reviews 34 24375389
2010 Promoter methylation of p16, Runx3, DAPK and CHFR genes is frequent in gastric carcinoma. Tumori 34 21302620
2008 Loss of CHFR in human mammary epithelial cells causes genomic instability by disrupting the mitotic spindle assembly checkpoint. Neoplasia (New York, N.Y.) 34 18592005
2010 First-line therapy and methylation status of CHFR in serum influence outcome to chemotherapy versus EGFR tyrosine kinase inhibitors as second-line therapy in stage IV non-small-cell lung cancer patients. Lung cancer (Amsterdam, Netherlands) 33 20705357
2018 The inhibition of UBC13 expression and blockage of the DNMT1-CHFR-Aurora A pathway contribute to paclitaxel resistance in ovarian cancer. Cell death & disease 32 29367628
2007 Relationship of aberrant DNA hypermethylation of CHFR with sensitivity to taxanes in endometrial cancer. Oncology reports 32 17143476
2004 CHFR-associated early G2/M checkpoint defects in breast cancer cells. Molecular carcinogenesis 32 14694445
2009 CHFR, a potential tumor suppressor, downregulates interleukin-8 through the inhibition of NF-kappaB. Oncogene 30 19448676
2007 Epigenetic inactivation of the CHFR gene in cervical cancer contributes to sensitivity to taxanes. International journal of oncology 30 17786301
2003 Chfr inactivation is not associated to chromosomal instability in colon cancers. Oncogene 30 14654793
2023 Ubiquitin ligase CHFR mediated degradation of VE-cadherin through ubiquitylation disrupts endothelial adherens junctions. Nature communications 29 37852964
2011 Association of CHFR promoter methylation with disease recurrence in locally advanced colon cancer. Clinical cancer research : an official journal of the American Association for Cancer Research 29 21551253
2004 PML bodies control the nuclear dynamics and function of the CHFR mitotic checkpoint protein. Nature structural & molecular biology 28 15467728
2015 Methylation of CHFR sensitizes esophageal squamous cell cancer to docetaxel and paclitaxel. Genes & cancer 27 25821560
2009 Functional interaction between Chfr and Kif22 controls genomic stability. The Journal of biological chemistry 27 19321445
2009 Deficiencies in Chfr and Mlh1 synergistically enhance tumor susceptibility in mice. The Journal of clinical investigation 27 19690386
2010 Epigenetic and genetic silencing of CHFR in esophageal adenocarcinomas. Cancer 25 20564104
2007 Yeast Chfr homologs retard cell cycle at G1 and G2/M via Ubc4 and Ubc13/Mms2-dependent ubiquitination. Cell cycle (Georgetown, Tex.) 25 18202552
2008 The anti-proliferative effects of the CHFR depend on the forkhead associated domain, but not E3 ligase activity mediated by ring finger domain. PloS one 24 18335050
2016 CHFR negatively regulates SIRT1 activity upon oxidative stress. Scientific reports 22 27883020
2011 CHFR hypermethylation and EGFR mutation are mutually exclusive and exhibit contrastive clinical backgrounds and outcomes in non-small cell lung cancer. International journal of cancer 22 20473935
2022 Circ_CHFR Promotes Platelet-Derived Growth Factor-BB-Induced Proliferation, Invasion, and Migration in Vascular Smooth Muscle Cells via the miR-149-5p/NRP2 Axis. Journal of cardiovascular pharmacology 21 33990513
2021 CHFR regulates chemoresistance in triple-negative breast cancer through destabilizing ZEB1. Cell death & disease 20 34462429
2006 Aberrant expression of CHFR in malignant peripheral nerve sheath tumors. Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc 20 16554732
2011 CHFR suppression by hypermethylation sensitizes endometrial cancer cells to paclitaxel. International journal of gynecological cancer : official journal of the International Gynecological Cancer Society 19 21792009
2006 DNA methylation of CHFR is not a predictor of the response to docetaxel and paclitaxel in advanced and recurrent gastric cancer. Anticancer research 19 16475678
2013 CHFR silencing or microsatellite instability is associated with increased antitumor activity of docetaxel or gemcitabine in colorectal cancer. International journal of cancer 18 23873170
2008 CHFR expression is preferentially impaired in smoking-related squamous cell carcinoma of the lung, and the diminished expression significantly harms outcomes. International journal of cancer 18 18623126
2004 Promoter hypermethylation and silencing of CHFR mitotic stress checkpoint gene in human gastric cancers. Oncology reports 18 15201973
2004 Hypermethylation of Chfr and hMLH1 in gastric noninvasive and early invasive neoplasias. Virchows Archiv : an international journal of pathology 18 15735977
2021 Vaccinia-related kinase 2 drives pancreatic cancer progression by protecting Plk1 from Chfr-mediated degradation. Oncogene 17 34140642
2006 Aberrant methylation of the CHFR gene in digestive tract cancer. Anticancer research 17 16827108
2017 Sustained expression of miR-26a promotes chromosomal instability and tumorigenesis through regulation of CHFR. Nucleic acids research 16 28126920
2010 Pathobiologic implications of methylation and expression status of Runx3 and CHFR genes in gastric cancer. Medical oncology (Northwood, London, England) 16 20300977
2005 Aberrant methylation of the CHFR gene in advanced hepatocellular carcinoma. Hepato-gastroenterology 16 16334792
2015 CHFR methylation strongly correlates with methylation of DNA damage repair and apoptotic pathway genes in non-small cell lung cancer. Discovery medicine 15 25828518
2010 CHFR functions as a ubiquitin ligase for HLTF to regulate its stability and functions. Biochemical and biophysical research communications 15 20388495
2008 Mechanism and pathobiologic implications of CHFR promoter methylation in gastric carcinoma. World journal of gastroenterology 15 18763281
2015 Small molecule inhibition of the CHFR-PARP1 interaction as novel approach to overcome intrinsic taxane resistance in cancer. Oncotarget 14 26356822
2009 Aberrant promoter hypermethylation of the CHFR gene in oral squamous cell carcinomas. Oncology reports 14 19787237
2006 Aberrant hypermethylation of the promoter region of the CHFR gene is rare in primary breast cancer. Breast cancer research and treatment 14 16502017
2006 Aberrant methylation of the CHFR gene is frequently detected in non-invasive colorectal cancer. Anticancer research 14 17201143
2012 Aggregated low-density lipoprotein induces LRP1 stabilization through E3 ubiquitin ligase CHFR downregulation in human vascular smooth muscle cells. Arteriosclerosis, thrombosis, and vascular biology 13 23241409
2021 Kaposi's sarcoma-associated herpesvirus processivity factor (PF-8) recruits cellular E3 ubiquitin ligase CHFR to promote PARP1 degradation and lytic replication. PLoS pathogens 12 33508027
2019 CHFR promotes the migration of human gastric cancer cells by inducing epithelial-to-mesenchymal transition in a HDAC1-dependent manner. OncoTargets and therapy 11 30799937
2018 Stabilization of HDAC1 via TCL1-pAKT-CHFR axis is a key element for NANOG-mediated multi-resistance and stem-like phenotype in immune-edited tumor cells. Biochemical and biophysical research communications 11 30060952
2018 Association of CHFR Promoter Methylation with Treatment Outcomes of Irinotecan-Based Chemotherapy in Metastatic Colorectal Cancer. Neoplasia (New York, N.Y.) 11 30562637
2012 SRG3/mBAF155 stabilizes the SWI/SNF-like BAF complex by blocking CHFR mediated ubiquitination and degradation of its major components. Biochemical and biophysical research communications 11 22285184
2022 Circ-CHFR modulates the proliferation, migration, and invasion of ox-LDL-induced human aorta vascular smooth muscle cells through the miR-214-3p/PAPPA axis. Clinical hemorheology and microcirculation 10 34842180
2011 The auto-ubiquitylation of E3 ubiquitin-protein ligase Chfr at G2 phase is required for accumulation of polo-like kinase 1 and mitotic entry in mammalian cells. The Journal of biological chemistry 10 21768102
2022 CHFR-Promoter-Methylation Status Is Predictive of Response to Irinotecan-based Systemic Chemotherapy in Advanced Colorectal Cancer. Anticancer research 9 35093868
2019 CHFR Promoter Hypermethylation Is Associated with Gastric Cancer and Plays a Protective Role in Gastric Cancer Process. Journal of Cancer 9 30854101
2014 Molecular characteristics of non-small cell lung cancer with reduced CHFR expression in The Cancer Genome Atlas (TCGA) project. Respiratory medicine 9 25477232
2010 Alternative efficacy-predicting markers for paclitaxel instead of CHFR in non-small-cell lung cancer. Cancer biology & therapy 9 20855974
2009 Nuclear localization of Chfr is crucial for its checkpoint function. Molecules and cells 9 19326084
2021 CHFR-mediated degradation of RNF126 confers sensitivity to PARP inhibitors in triple-negative breast cancer cells. Biochemical and biophysical research communications 8 34388456
2020 Analysis of the methylation of CpG islands in the CDO1, TAC1 and CHFR genes in pancreatic ductal cancer. Oncology letters 8 32194717
2019 Silencing of CHFR Sensitizes Gastric Carcinoma to PARP Inhibitor Treatment. Translational oncology 8 31812083
2016 Association between CHFR gene hypermethylation and gastric cancer risk: a meta-analysis. OncoTargets and therapy 8 27994471
2015 CHFR hypermethylation, a frequent event in acute myeloid leukemia, is independently associated with an adverse outcome. Genes, chromosomes & cancer 8 26542416
2021 circ_CHFR regulates ox-LDL-mediated cell proliferation, apoptosis, and EndoMT by miR-15a-5p/EGFR axis in human brain microvessel endothelial cells. Open life sciences 7 34676300
2008 Coding region polymorphisms in the CHFR mitotic stress checkpoint gene are associated with colorectal cancer risk. Cancer letters 7 18079053
2017 Upregulation of the checkpoint protein CHFR is associated with tumor suppression in pancreatic cancers. Oncology letters 5 29344247
2011 CHFR binds to and regulates MAD2 in the spindle checkpoint through its cysteine-rich domain. Biochemical and biophysical research communications 5 21575600
2010 Promoter methylation of CHFR gene in gastric carcinoma tissues detected using two methods. Chinese journal of cancer 5 20109344
2009 CHFR promoter hypermethylation and reduced CHFR mRNA expression in ovarian cancer. The International journal of biological markers 5 19634111

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