Affinage

ZNF224

Zinc finger protein 224 · UniProt Q9NZL3

Length
707 aa
Mass
82.3 kDa
Annotated
2026-06-11
20 papers in source corpus 16 papers cited in narrative 16 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

ZNF224 is a KRAB-domain Krüppel-like zinc finger protein that functions as a sequence-specific transcription factor, binding a 5'-CAGC-3' consensus motif and acting predominantly as a repressor while adopting context-dependent activating or modulatory roles depending on cell type and partner (PMID:16150558, PMID:27105517). Its core repressive mechanism relies on the KRAB A domain, which recruits the co-repressor KAP-1 and histone deacetylase activity to silence target promoters such as aldolase A (PMID:16150558). ZNF224 also assembles a repressor complex with the arginine methyltransferase PRMT5, directing symmetric dimethylation of histone H4 arginine 3 at target promoters in a cell-cycle-dependent manner (PMID:19741270); documented direct repression targets additionally include the mitochondrial citrate carrier CIC (PMID:19505435). Beyond autonomous repression, ZNF224 acts as a co-regulator of WT1, enhancing WT1-driven activation of proapoptotic genes while counteracting WT1-mediated repression of others such as IRF8 (PMID:23362234, PMID:26563595). In chronic myeloid leukemia ZNF224 is a tumor-suppressive node repressing c-Myc and AXL, and its expression is suppressed by BCR-ABL/WT1 signaling but restored by kinase inhibitors to promote apoptosis (PMID:26320177, PMID:29423056, PMID:30176265). In other contexts ZNF224 supports survival and oncogenic phenotypes: it activates miR-663a to lower p53/p21 (PMID:27105517), promotes NF-κB-dependent survival in CLL (PMID:36425656), potentiates TGF-β/Smad-driven EMT in melanoma (PMID:34181020), and drives AKT-dependent cytosolic retention of p21 (PMID:40321146). Its activity and stability are further shaped by partner binding, including DEPDC1, which sequesters ZNF224 to repress the NF-κB inhibitor A20 (PMID:20587513), and MED28, which binds the KRAB domain and protects ZNF224 from degradation (PMID:29435049).

Mechanistic history

Synthesis pass · year-by-year structured walk · 16 steps
  1. 2005 High

    Established ZNF224 as a bona fide KRAB-zinc finger repressor by showing it occupies a target promoter in vivo and silences it through the canonical KRAB-A/KAP-1/HDAC axis.

    Evidence ChIP, reporter assays, KRAB domain mutagenesis, and HDAC inhibitor (TSA) treatment on the aldolase A promoter

    PMID:16150558

    Open questions at the time
    • Did not define the full genomic target repertoire
    • Recruitment of specific HDAC enzymes not identified
  2. 2007 Medium

    Showed that alternative splicing generates the ZNF255 isoform with distinct subnuclear/cytoplasmic localization and weaker repressive capacity, linking subcellular distribution to function.

    Evidence Northern blot, GFP-tagged protein imaging, and ChIP comparing ZNF224 and ZNF255

    PMID:17900823

    Open questions at the time
    • Functional role of cytoplasmic/nucleolar ZNF255 undefined
    • Single lab, no endogenous protein detection
  3. 2009 High

    Defined the enzymatic effector arm of ZNF224 repression by identifying PRMT5 as a complex partner that deposits the repressive H4R3 symmetric dimethyl mark in a cell-cycle-dependent manner.

    Evidence Reciprocal Co-IP, ChIP, PRMT5 RNAi de-repression, and histone methylation assay at the aldolase A promoter

    PMID:19741270

    Open questions at the time
    • Whether PRMT5 recruitment generalizes beyond aldolase A unknown
    • Order of KAP-1 vs PRMT5 recruitment not resolved
  4. 2009 Medium

    Extended the repressive target set by mapping a discrete silencer element bound by ZNF224 in the CIC promoter and showing bidirectional control of CIC expression.

    Evidence DNA affinity purification, luciferase reporters, and ZNF224 overexpression/siRNA with RT-PCR and Western blot

    PMID:19505435

    Open questions at the time
    • Direct in vivo promoter occupancy by ChIP not shown
    • Co-repressor requirement at CIC not tested
  5. 2010 Medium

    Revealed that ZNF224 is not a constitutive repressor but a partner-dependent co-regulator, acting as a WT1(−KTS) co-activator, recasting it as a context-switchable transcription factor.

    Evidence Co-IP, subcellular fractionation/immunofluorescence, and reporter assays with WT1 isoforms

    PMID:20591825

    Open questions at the time
    • Structural basis of the activating vs repressive switch unknown
    • Single lab
  6. 2010 High

    Demonstrated a non-promoter sequestration mechanism whereby DEPDC1 binding restrains ZNF224 from repressing the NF-κB inhibitor A20, linking ZNF224 partner availability to survival signaling.

    Evidence Reciprocal Co-IP, colocalization, cell-permeable peptide disruption, and in vitro/xenograft apoptosis assays in bladder cancer

    PMID:20587513

    Open questions at the time
    • Direct ZNF224 binding to the A20 promoter not mapped
    • Generality beyond bladder cancer untested
  7. 2013 Medium

    Showed ZNF224 differentially co-regulates WT1 targets in leukemia—activating proapoptotic and suppressing antiapoptotic genes—and that chemotherapy induces ZNF224 to enhance apoptosis.

    Evidence Reporter assays, Co-IP, ChIP, siRNA, and apoptosis assays with cytarabine in K562 cells

    PMID:23362234

    Open questions at the time
    • Determinant of activating vs repressive outcome on individual WT1 targets unresolved
    • Single lab
  8. 2015 Medium

    Mapped a regulatory feedback whereby BCR-ABL, via WT1, transcriptionally represses ZNF224, establishing ZNF224 silencing as a route by which the oncogene blunts apoptosis.

    Evidence Promoter reporter assays, expression analysis in BCR-ABL+ lines and primary CML, and TKI treatment

    PMID:26320177

    Open questions at the time
    • Direct WT1 occupancy of the ZNF224 promoter in vivo not shown
    • Single lab
  9. 2015 Medium

    Identified IRF8 as a target where ZNF224 counteracts WT1-mediated repression, reinforcing its role as a WT1 co-regulator in leukemia differentiation/apoptosis.

    Evidence Nuclear Co-IP, ChIP, shRNA knockdown, and reporter assays in K562 cells

    PMID:26563595

    Open questions at the time
    • Mechanism by which ZNF224 overrides WT1 repression unclear
    • Single lab
  10. 2016 High

    Defined the ZNF224 DNA-binding consensus (5'-CAGC-3') genome-wide and uncovered an activating, pro-survival arm via miR-663a induction that lowers p53 and p21.

    Evidence ChIP-seq, SPR binding, reporter assays, and miR-663a antagonist rescue

    PMID:27105517

    Open questions at the time
    • How the same factor activates miR-663a yet represses other genes mechanistically unresolved
    • Cofactors driving activation not identified
  11. 2017 Medium

    Established ZNF224 as a tumor-suppressive repressor of c-Myc whose drug-induced expression promotes apoptosis, defining a ZNF224/c-Myc axis governing imatinib response.

    Evidence c-Myc promoter reporters, gain/loss-of-function, and imatinib/AG490 treatment with apoptosis assays in CML

    PMID:29423056

    Open questions at the time
    • Direct c-Myc promoter occupancy and co-repressor use not fully detailed
    • Single lab
  12. 2017 Medium

    Showed MED28 binds the ZNF224 KRAB domain and stabilizes ZNF224 against degradation, identifying post-translational control of ZNF224 abundance as a regulatory layer.

    Evidence Reciprocal Co-IP, SPR, BiFC, domain mapping, and colony formation in MCF-7 cells

    PMID:29435049

    Open questions at the time
    • Degradation pathway (E3 ligase, ubiquitination) not identified
    • Effect on ZNF224 transcriptional targets not assessed
  13. 2018 Medium

    Identified AXL as an additional ZNF224 repression target in CML, linking ZNF224 loss to imatinib resistance and its restoration to resensitization.

    Evidence AXL promoter reporters, ChIP, gain/loss-of-function, and drug sensitivity assays

    PMID:30176265

    Open questions at the time
    • Co-repressor machinery at AXL not characterized
    • Single lab
  14. 2021 Medium

    Revealed an oncogenic role in melanoma where TGF-β-induced ZNF224 potentiates Smad-driven EMT transcription and feeds back to upregulate TGF-β and its receptors.

    Evidence Gain/loss-of-function, TGF-β stimulation, EMT marker analysis, invasion/proliferation, and reporter assays

    PMID:34181020

    Open questions at the time
    • Direct ZNF224 targets within the EMT program not mapped
    • Single lab
  15. 2022 Medium

    Demonstrated ZNF224 promotes CLL survival through NF-κB pathway modulation, with knockdown raising apoptosis and reducing proliferation in primary patient cells.

    Evidence RNAi with NF-κB activity, apoptosis, and proliferation assays in primary CLL PBMCs

    PMID:36425656

    Open questions at the time
    • Direct transcriptional targets connecting ZNF224 to NF-κB undefined
    • Single lab
  16. 2025 Medium

    Showed ZNF224 co-opts p21 oncogenically in melanoma by transcriptionally upregulating p21 in a p53-dependent manner and promoting AKT-driven cytosolic retention to block apoptosis.

    Evidence p21 promoter reporters, p53-dependency and AKT-inhibition experiments, subcellular fractionation, and apoptosis/proliferation assays

    PMID:40321146

    Open questions at the time
    • Direct ZNF224 binding to the p21 promoter versus indirect effect not distinguished
    • Single lab

Open questions

Synthesis pass · forward-looking unresolved questions
  • A unifying mechanistic determinant of when ZNF224 acts as a KAP-1/PRMT5-dependent repressor versus a WT1/Smad co-activator across cell types remains unresolved.
  • No structural model of partner-dependent activity switching
  • Genome-wide direct targets in non-leukemic contexts uncharacterized
  • Post-translational code governing repression vs activation unknown

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0140110 transcription regulator activity 6 GO:0003677 DNA binding 3
Localization
GO:0005634 nucleus 3 GO:0005730 nucleolus 1
Pathway
R-HSA-74160 Gene expression (Transcription) 5 R-HSA-5357801 Programmed Cell Death 4 R-HSA-4839726 Chromatin organization 2
Partners
DEPDC1KAP1MED28PRMT5WT1

Evidence

Reading pass · 16 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2005 ZNF224 binds in vivo to the distal promoter of the aldolase A gene and represses its transcription; this repression requires the 45-amino acid KRAB A domain and its interaction with the KAP-1 co-repressor, and also requires histone deacetylases (demonstrated by TSA sensitivity). Chromatin immunoprecipitation (ChIP), transient transfection/reporter assays, co-transfection with HDAC inhibitor trichostatin A, KRAB domain mutagenesis Gene High 16150558
2009 ZNF224 physically associates with the arginine methyltransferase PRMT5; PRMT5 is recruited to the L-type aldolase A promoter as part of the ZNF224 repressor complex, leading to symmetric dimethylation of arginine 3 of histone H4. RNAi knockdown of PRMT5 de-represses aldolase A transcription. The ZNF224–PRMT5 repressor complex occupies the promoter in a cell-cycle-dependent manner. Co-immunoprecipitation, chromatin immunoprecipitation (ChIP), RNA interference, histone methylation assay The Journal of biological chemistry High 19741270
2009 ZNF224 binds a 26-bp silencer element (-595/-569) in the mitochondrial citrate carrier (CIC) promoter and represses CIC transcription; overexpression of ZNF224 decreases CIC reporter and endogenous transcript/protein, while ZNF224 knockdown activates CIC transcription. DNA affinity purification/protein identification, reporter (luciferase) assays, ZNF224 overexpression and siRNA knockdown, RT-PCR and Western blot for CIC Biochemical and biophysical research communications Medium 19505435
2010 ZNF224 interacts with DEPDC1 (cancer-testis antigen) in bladder cancer cells, as shown by co-immunoprecipitation and colocalization. Disruption of the DEPDC1–ZNF224 complex with a cell-permeable peptide de-represses A20 transcription (an NF-κB inhibitor), triggering apoptosis in vitro and in vivo. Co-immunoprecipitation, immunocytochemistry/colocalization, cell-permeable peptide interference, luciferase/transcriptional assays, in vitro and xenograft apoptosis assays Cancer research High 20587513
2010 ZNF224 (nuclear isoform) physically interacts with the WT1(−KTS) isoform in the nucleus, acting as a co-activator that enhances WT1-mediated transcriptional activation (e.g., of the vitamin D receptor promoter). ZNF255 (cytoplasmic/nucleolar isoform) interacts with WT1(+KTS) in a different subcellular compartment and is involved in RNA processing rather than transcription. Co-immunoprecipitation, subcellular fractionation/immunofluorescence, reporter (luciferase) transcriptional assays Human molecular genetics Medium 20591825
2007 ZNF224 is homogeneously distributed in the nucleus, whereas its isoform ZNF255 (alternatively spliced from the same gene) localizes to subnuclear structures (nucleoli) and also to the cytoplasm; ZNF224 represses aldolase A NRE more efficiently than ZNF255, correlating with their distinct subcellular distributions. Northern blot, PCR, transient transfection with GFP-tagged proteins (immunofluorescence), ChIP Gene Medium 17900823
2013 In CML K562 cells, ZNF224 acts as a co-activator of WT1 on proapoptotic gene promoters while suppressing WT1-mediated transactivation of antiapoptotic genes; cytarabine (ara-C) induces ZNF224 expression, and this induction enhances the apoptotic response to ara-C. Reporter (luciferase) assays, co-immunoprecipitation, ChIP, siRNA knockdown, apoptosis assays Human molecular genetics Medium 23362234
2015 BCR-ABL oncogene signaling downregulates ZNF224 gene expression in CML; WT1 (whose expression is positively driven by BCR-ABL) transcriptionally represses the ZNF224 promoter, providing a pathway by which BCR-ABL suppresses apoptosis through ZNF224 repression. RT-PCR and Western blot in BCR-ABL+ cell lines and primary CML samples, reporter assays for ZNF224 promoter, imatinib/TKI treatment experiments Oncotarget Medium 26320177
2015 ZNF224 is recruited by WT1 to the IRF8 promoter and counteracts WT1-mediated repression of IRF8 transcription; ZNF224 knockdown suppresses both basal and cytarabine-induced IRF8 expression in K562 cells. Co-immunoprecipitation (nuclear), ChIP, shRNA knockdown, luciferase reporter assays Leukemia research Medium 26563595
2016 ZNF224 binds a consensus 5'-CAGC-3' DNA motif (identified by ChIP-seq) and transcriptionally activates miR-663a by binding its promoter; miR-663a in turn binds the 3' UTR of p53 and p21 to reduce their expression, thereby promoting cell survival and apoptosis resistance. ChIP-sequencing, ELISA, surface plasmon resonance (SPR), qPCR, luciferase reporter assay, miR-663a antagonist rescue experiments Oncotarget High 27105517
2017 ZNF224 is a transcriptional repressor of c-Myc in CML cells; imatinib and the JAK2 inhibitor AG490 induce ZNF224 expression, which then represses c-MYC transcription, leading to apoptosis. This identifies the ZNF224/c-Myc axis as a determinant of imatinib responsiveness. Reporter (luciferase) assays for c-Myc promoter, ZNF224 overexpression/knockdown, RT-PCR and Western blot, drug treatment experiments (imatinib, AG490), apoptosis assays Oncotarget Medium 29423056
2017 MED28 physically interacts with ZNF224 via the KRAB domain of ZNF224 and the MED domain of MED28, in the nucleus; MED28 overexpression stabilizes ZNF224 protein by inhibiting its camptothecin (CPT)-induced degradation, resulting in increased colony formation of MCF-7 breast cancer cells. Co-immunoprecipitation, surface plasmon resonance (SPR), bimolecular fluorescence complementation (BiFC), domain mapping, colony formation assay Oncology letters Medium 29435049
2018 ZNF224 acts as a transcriptional repressor of AXL (receptor tyrosine kinase) in CML cells; ZNF224 overexpression suppresses AXL expression and partly restores imatinib sensitivity in imatinib-resistant CML cells. Reporter assays (AXL promoter), ChIP, ZNF224 overexpression/knockdown, Western blot, drug sensitivity assays Biochimie Medium 30176265
2021 ZNF224 expression is induced by TGF-β in melanoma cells and mediates TGF-β pro-oncogenic signaling; ZNF224 potentiates TGF-β/Smad-driven transcription of EMT target genes and positively modulates expression of TGF-β itself and its receptors TβR1 and TβR2, establishing a positive regulatory loop that promotes EMT and invasiveness. ZNF224 overexpression and knockdown, TGF-β stimulation, RT-PCR/Western blot for EMT markers and TGF-β pathway components, cell invasion/proliferation assays, reporter assays Human molecular genetics Medium 34181020
2022 ZNF224 knockdown reduces NF-κB pathway activity in CLL cells, demonstrating that ZNF224 promotes CLL cell survival through NF-κB pathway modulation; ZNF224 silencing raises both spontaneous and drug-induced apoptosis and inhibits proliferation in primary CLL cells. RNA interference (ZNF224 knockdown), NF-κB reporter/activity assay, apoptosis assays, proliferation assays in primary CLL patient PBMCs Frontiers in molecular biosciences Medium 36425656
2025 In melanoma cells, ZNF224 overexpression transcriptionally upregulates p21(CIP1/WAF1) in a p53-dependent manner and enhances AKT-triggered cytosolic retention of p21, inhibiting apoptosis and promoting cell proliferation; this reveals a mechanism by which ZNF224 co-opts p21's oncogenic function downstream of p53 and AKT signaling. ZNF224 overexpression/knockdown, p21 promoter reporter assays, p53 dependency experiments, AKT pathway inhibition, subcellular fractionation, apoptosis and proliferation assays, transcriptomic data analysis The FEBS journal Medium 40321146

Source papers

Stage 0 corpus · 20 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2010 Cell-permeable peptide DEPDC1-ZNF224 interferes with transcriptional repression and oncogenicity in bladder cancer cells. Cancer research 86 20587513
2005 The Krüppel-like zinc-finger protein ZNF224 represses aldolase A gene transcription by interacting with the KAP-1 co-repressor protein. Gene 33 16150558
2016 ZNF224, Krüppel like zinc finger protein, induces cell growth and apoptosis-resistance by down-regulation of p21 and p53 via miR-663a. Oncotarget 31 27105517
2009 The Kruppel-like zinc finger protein ZNF224 recruits the arginine methyltransferase PRMT5 on the transcriptional repressor complex of the aldolase A gene. The Journal of biological chemistry 31 19741270
2010 Biochemical and functional interaction between ZNF224 and ZNF255, two members of the Kruppel-like zinc-finger protein family and WT1 protein isoforms. Human molecular genetics 26 20591825
2009 Transcription of the mitochondrial citrate carrier gene: identification of a silencer and its binding protein ZNF224. Biochemical and biophysical research communications 24 19505435
2013 Role of WT1-ZNF224 interaction in the expression of apoptosis-regulating genes. Human molecular genetics 22 23362234
2015 WT1-mediated repression of the proapoptotic transcription factor ZNF224 is triggered by the BCR-ABL oncogene. Oncotarget 19 26320177
2021 ZNF224 is a mediator of TGF-β pro-oncogenic function in melanoma. Human molecular genetics 16 34181020
2017 Role of ZNF224 in c-Myc repression and imatinib responsiveness in chronic myeloid leukemia. Oncotarget 15 29423056
2015 The hematopoietic tumor suppressor interferon regulatory factor 8 (IRF8) is upregulated by the antimetabolite cytarabine in leukemic cells involving the zinc finger protein ZNF224, acting as a cofactor of the Wilms' tumor gene 1 (WT1) protein. Leukemia research 15 26563595
2017 Role of ZNF224 in cell growth and chemoresistance of chronic lymphocitic leukemia. Human molecular genetics 14 28040726
2010 ZNF224: Structure and role of a multifunctional KRAB-ZFP protein. The international journal of biochemistry & cell biology 14 21187159
2007 Differential expression and cellular localization of ZNF224 and ZNF255, two isoforms of the Krüppel-like zinc-finger protein family. Gene 14 17900823
2018 ZNF224 is a transcriptional repressor of AXL in chronic myeloid leukemia cells. Biochimie 12 30176265
2016 The Complex Role of the ZNF224 Transcription Factor in Cancer. Advances in protein chemistry and structural biology 12 28215224
2017 MED28 increases the colony-forming ability of breast cancer cells by stabilizing the ZNF224 protein upon DNA damage. Oncology letters 9 29435049
2022 Biological relevance of ZNF224 expression in chronic lymphocytic leukemia and its implication IN NF-kB pathway regulation. Frontiers in molecular biosciences 7 36425656
2021 ZNF224 Protein: Multifaceted Functions Based on Its Molecular Partners. Molecules (Basel, Switzerland) 6 34684876
2025 ZNF224 enhances the oncogenic function of p21 via p53 and AKT pathways in melanoma. The FEBS journal 1 40321146

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