| 1991 |
ATBF1 (ZFHX3) was identified as a large DNA-binding protein (306 kDa) containing four homeodomains and 17 zinc finger motifs that binds to an AT-rich motif in the human alpha-fetoprotein (AFP) gene enhancer, establishing its identity as a transcriptional regulatory protein. |
cDNA cloning and sequencing; DNA-binding assays |
Molecular and cellular biology |
Medium |
1719379
|
| 1994 |
ATBF1 selectively suppresses AFP enhancer and promoter activities by binding to AT-rich elements (AT motifs); suppression was abrogated by site-specific mutagenesis of the AT motif. ATBF1 competes with HNF1 for the same binding site, providing a mechanism for selective AFP (but not albumin) repression. |
Gel retardation (EMSA), site-directed mutagenesis, cotransfection/CAT reporter assays |
Molecular and cellular biology |
High |
7507206
|
| 1995 |
ATBF1 exists as two isoforms generated by alternative promoter usage and splicing: ATBF1-B (306 kDa, 4 homeodomains, 18 zinc fingers) and ATBF1-A (404 kDa, with an additional 920-amino acid N-terminal extension). ATBF1-A expression is driven by a neuronal differentiation-dependent promoter activated during retinoic acid-induced neuronal differentiation of P19 and NT2/D1 cells. |
cDNA cloning, Northern blot, RNase protection assay, transient transfection/CAT reporter assay |
The Journal of biological chemistry |
High |
7592926
|
| 1999 |
ATBF1 physically interacts with v-Myb and c-Myb proteins; the interaction requires amino acids 2484–2520 of ATBF1 and 279–300 of v-Myb (plus Leu325/Leu332 of the Myb leucine zipper). ATBF1 forms in vivo complexes with v-Myb and inhibits v-Myb transcriptional activity in co-transfection experiments. |
Yeast two-hybrid, in vitro binding/deletion mutagenesis, co-transfection luciferase assay, co-immunoprecipitation |
The Journal of biological chemistry |
High |
10318867
|
| 2001 |
ATBF1-A isoform inhibits myogenic differentiation of C2C12 cells by repressing MyoD and myogenin and elevating Id3 and cyclin D1; the fourth homeodomain of ATBF1-A binds an AT-rich element adjacent to the E1 E-box of the MRF4 promoter to mediate transcriptional repression. The ATBF1-A-specific N-terminal region acts as a general transcriptional repressor. Conversely, ATBF1-B accelerates myogenic differentiation. |
Forced expression/transfection, Western blot, promoter-reporter assay, EMSA with defined homeodomain construct |
The Journal of biological chemistry |
High |
11312261
|
| 2001 |
ATBF1-A directly binds an AT motif in the aminopeptidase N (APN) gene promoter and suppresses APN transcription in the crypt-villus axis of the small intestine; deletion of the AT motif abolishes suppression. ATBF1-A expression decreases upon butyrate-induced enterocyte differentiation. |
In situ hybridization, transient transfection/dual luciferase reporter assay with AT-motif deletion construct, RT-PCR |
Biochemical and biophysical research communications |
Medium |
10623580
|
| 2001 |
ATBF1-A protein possesses an intrinsic DNA/RNA-dependent ATPase activity; a 263-amino acid segment (AHZ) containing the ATPase A-motif, homeodomain IV, and zinc finger 21 hydrolyzes ATP in a DNA/RNA-dependent manner. Site-directed mutagenesis of homeodomain IV and zinc finger 21 (but not the A-motif alone) abolished both DNA/RNA binding and ~80–90% of ATPase activity, establishing these domains as essential for enzymatic function. |
In vitro ATPase assay with recombinant GST-fusion protein, site-directed mutagenesis |
Biochimica et biophysica acta |
High |
11755205
|
| 2004 |
ATBF1 interacts with PIAS3 (identified by yeast two-hybrid) and forms in vivo complexes with PIAS3 in HepG2 cells; ATBF1 alone has no effect on STAT3 signaling but synergistically enhances PIAS3-mediated inhibition of IL-6-activated STAT3, suppressing IL-6-mediated cellular responses. |
Yeast two-hybrid, co-immunoprecipitation, luciferase reporter assay (STAT3-responsive) |
Biochemical and biophysical research communications |
Medium |
14715251
|
| 2004 |
ATBF1 directly interacts with p53 (co-immunoprecipitation) and stimulates p21(Waf1/Cip1) promoter activity; MMC treatment and ATBF1 overexpression synergistically activated p21 promoter activity up to 7-fold, placing ATBF1 in a p53-p21 cell cycle regulatory pathway. |
Immunoprecipitation, dual luciferase promoter-reporter assay |
Microbiology and immunology |
Medium |
14978340
|
| 2005 |
ATBF1 overexpression in Neuro 2A cells causes cell cycle arrest and nuclear localization; in P19 cells, ATBF1 is cytoplasmic in floating/nonadherent conditions (due to CRM1-dependent nuclear export) but translocates to the nucleus and arrests the cell cycle upon cell-matrix adhesion. Nuclear localization (and hence cell cycle arrest) is suppressed by caffeine (an ATM inhibitor), and cytoplasmic retention requires CRM1-dependent export. |
Overexpression, cell cycle analysis, pharmacological inhibition (caffeine, leptomycin B), fluorescence microscopy |
Development (Cambridge, England) |
Medium |
16251211
|
| 2007 |
Nuclear ATBF1 suppresses MUC5AC gene expression in gastric cancer by binding to an AT motif-like element in the MUC5AC promoter; chromatin immunoprecipitation confirmed ATBF1 occupancy at this element, and deletion of the AT motif abolished ATBF1-mediated suppression in reporter assays. |
Dual luciferase reporter assay, chromatin immunoprecipitation (ChIP), overexpression with endogenous MUC5AC protein analysis, immunohistochemistry |
International journal of cancer |
High |
17330845
|
| 2008 |
Atbf1 is required for early activation of the Pit1 gene in the developing pituitary; Atbf1 binds to and activates one of two critical distant enhancers of the Pit1 locus, as demonstrated by in vivo enhancer mapping combined with proteomics (mass spectrometry) identification and genetic/in vitro validation. |
In vivo enhancer mapping, proteomics/mass spectrometry, genetic analysis, in vitro transcription assay |
Proceedings of the National Academy of Sciences of the United States of America |
High |
18272476
|
| 2010 |
ZFHX3 (ATBF1) induces expression of PDGFRB and thereby activates ATM in the cytoplasm under oxidative stress (but not genotoxic stress), defining a novel signaling axis: ATM→CREB→ZFHX3→PDGFRB→cytoplasmic ATM activation for neuronal protection. PDGFRB inhibition with AG1433 suppressed oxidative-stress-induced ATM activation but not X-ray-induced ATM activation. |
Pharmacological inhibition (AG1433), immunofluorescence/subcellular fractionation, reporter/expression assays in primary cerebellar neurons and cell lines |
Disease models & mechanisms |
Medium |
20876357
|
| 2010 |
ATBF1 inhibits estrogen receptor (ER)-mediated transcription and cell proliferation in ER-positive breast cancer cells by physically interacting with ER (in vitro and in vivo immunoprecipitation) and selectively competing with the coactivator AIB1 (but not GRIP1 or SRC1) for ER binding. |
In vitro and in vivo co-immunoprecipitation, competition binding assays, luciferase reporter assay, cell proliferation assay |
The Journal of biological chemistry |
High |
20720010
|
| 2010 |
ATBF1 physically associates with RUNX3 (co-immunoprecipitation) and both proteins translocate together to the nucleus in response to TGF-β1 stimulation in gastric cancer cells. ATBF1 and RUNX3 synergistically upregulate p21(Waf1/Cip1) promoter activity. |
Co-immunoprecipitation, confocal laser scanning microscopy (subcellular localization), dual luciferase reporter assay, immunohistochemistry |
Biochemical and biophysical research communications |
Medium |
20599712
|
| 2011 |
Estrogen upregulates ATBF1 transcription via direct binding of ERα to a half-estrogen-responsive element in the ATBF1 promoter; at higher estrogen levels, ATBF1 protein is degraded via the proteasome system, creating an autoregulatory feedback loop between ATBF1 and estrogen-ERα signaling. |
Promoter-reporter assay, ChIP (ER binding to ATBF1 promoter), Western blot/protein stability assay, pharmacological proteasome inhibition |
The Journal of biological chemistry |
High |
21367855
|
| 2012 |
EFP (oestrogen-responsive finger protein) is the E3 ubiquitin ligase that mediates oestrogen-induced ATBF1 protein ubiquitination and proteasomal degradation; EFP knockdown increases ATBF1 protein levels, EFP overexpression decreases them, and EFP physically interacts with and ubiquitinates ATBF1. |
RNAi knockdown, overexpression, co-immunoprecipitation, ubiquitination assay, Western blot |
The Biochemical journal |
High |
22452784
|
| 2012 |
ATBF1 nuclear localization is defined by a 3-amino acid NLS motif (KRK2615-2617); ectopically expressed ATBF1 forms nuclear body-like dots that physically associate with PML nuclear bodies. ATBF1 is SUMOylated at multiple lysine residues (K2349, K2806, K3258) in a nuclear-specific manner. PIAS3, which directly interacts with ATBF1, paradoxically diminishes rather than enhances ATBF1 SUMOylation. |
Site-directed mutagenesis of NLS, fluorescence microscopy/co-localization with PML, SUMOylation assay, co-immunoprecipitation |
PloS one |
Medium |
24651376
|
| 2012 |
ATBF1 protein is phosphorylated at multiple serine residues in embryonic brain (Ser1600, Ser2634, Ser2795, Ser2804, Ser2900, Ser3431, Ser3613, Ser3697) and this phosphorylation protects ATBF1 from cleavage by calpain-1. Dephosphorylation with alkaline phosphatase sensitizes ATBF1 to calpain-1 digestion; calcineurin inhibition with FK506 enhances ATBF1 resistance to calpain-1. |
Mass spectrometry phosphopeptide identification, in vitro calpain-1 cleavage assay, alkaline phosphatase treatment, FK506 pharmacological treatment |
Biochemical and biophysical research communications |
High |
23022192
|
| 2012 |
Progesterone-PR signaling directly induces ATBF1 transcription; activated PR binds to the ATBF1 promoter (demonstrated by ChIP) and upregulates ATBF1 at both mRNA and protein levels. ATBF1 induction is required for Pg-promoted mammary progenitor cell expansion, as prevention of ATBF1 induction inhibited Pg-PR-driven colony formation and stem cell marker expression. |
ChIP (PR binding to ATBF1 promoter), promoter-reporter assay, RNAi knockdown, RT-PCR, Western blot, Matrigel colony assay |
Biochemical and biophysical research communications |
High |
23159610
|
| 2015 |
ZFHX3 regulates circadian transcription via direct interaction with AT motifs in target gene promoters in the SCN. A dominant missense mutation (Zfhx3^Sci) decreases ZFHX3's ability to activate AT motifs in vitro; in Zfhx3^Sci/+ SCN, neuropeptide gene expression (critical for intercellular signaling) is disturbed while core clock genes are largely unaffected. Lentiviral transduction showed ZFHX3-mediated AT motif activation is circadian with reduced amplitude in mutant SCN. |
Dominant missense mouse mutant, RNA sequencing, in vitro AT-motif reporter assay, lentiviral SCN transduction, behavioral circadian analysis |
Cell |
High |
26232227
|
| 2016 |
ZFHX3 knockdown in HL-1 atrial myocytes dysregulates calcium homeostasis (increased SR Ca2+ content, Ca2+ transient, and Ca2+ leak) and increases atrial arrhythmogenesis by altering expression of SERCA2a, ryanodine receptor, Kv1.4, Kv1.5, Kir3.4, and associated ionic currents. |
Stable shRNA knockdown, patch clamp, confocal fluorescence microscopy (calcium imaging), Western blot |
International journal of cardiology |
Medium |
26930642
|
| 2016 |
ZFHX3 maintains prolactin receptor (Prlr) expression and Prlr-Jak2-Stat5 signaling activity in mammary epithelial cells; Zfhx3 knockout in mouse mammary tissue interrupts lactogenesis, reduces β-casein expression, and causes failure of lactation with underdeveloped alveoli. |
Conditional knockout, Western blot (Prlr, Stat5 phosphorylation), RT-PCR, histology, HC11 cell knockdown model |
The Journal of biological chemistry |
High |
27129249
|
| 2017 |
Adult-specific inducible knockout of Zfhx3 in mice shortens circadian free-running period by >1 h and causes complete behavioral arrhythmicity in 30% of homozygous animals, confirming a sustained role for ZFHX3 in adult SCN circadian rhythmicity independent of developmental effects. |
Tamoxifen-inducible Cre-mediated conditional knockout (UBC-cre/ERT2), wheel-running circadian behavior analysis before and after excision in same animals |
Journal of biological rhythms |
High |
28816086
|
| 2019 |
ZFHX3 and ERβ physically interact with each other in prostate cancer cells; both occupy the same region of the MYC promoter, and loss of ZFHX3 prevents ERβ from repressing MYC transcription and inhibiting cell proliferation, establishing MYC repression as a mechanism for ZFHX3's tumor suppressor activity. |
Co-immunoprecipitation, ChIP (ZFHX3 and ERβ on MYC promoter), siRNA knockdown, cell proliferation assay, luciferase reporter assay |
Oncogenesis |
High |
30979864
|
| 2019 |
Zfhx3 is essential for progesterone/PR signaling-driven ductal side-branching and alveologenesis; postnatal Zfhx3 deletion attenuates Pg-PR-mediated branching morphogenesis. ZFHX3 physically associates with PR (co-IP), and Pg/PR signaling expands PR/Zfhx3-positive epithelial cells. Zfhx3 loss impairs PR-dependent gene transcription. |
Conditional knockout, co-immunoprecipitation (ZFHX3-PR), histology, RT-PCR, Western blot |
Journal of genetics and genomics |
High |
30954439
|
| 2019 |
ZFHX3 regulates calcium influx in mammary epithelial cells partly via transcriptional regulation of the TRPV6 calcium channel; ZFHX3 knockdown reduces TRPV6 expression (confirmed by luciferase reporter assay of TRPV6 promoter), and calcium entry in ZFHX3-KD cells is partially rescued by wild-type but not pore-dead TRPV6. |
siRNA knockdown, luciferase reporter assay (TRPV6 promoter), calcium imaging, rescue with TRPV6 mutants |
Biochemical and biophysical research communications |
Medium |
31519324
|
| 2020 |
SUMOylation of ZFHX3 at Lys-2806 (the major site) is mediated by E1 enzyme SAE1, E2 enzyme UBC9, and E3 ligase PIAS2; SUMO-specific peptidases SENP1 and SENP2 deSUMOylate ZFHX3. SUMOylation at Lys-2806 enhances ZFHX3 stability by interfering with ubiquitination/proteasomal degradation, and promotes ZFHX3-mediated cell proliferation and xenograft tumor growth. |
SUMOylation assay with identified enzymes, mutagenesis of SUMOylation sites, ubiquitination assay, protein stability analysis, cell proliferation/xenograft assay |
The Journal of biological chemistry |
High |
32249212
|
| 2020 |
ZFHX3 physically associates with HIF1A and is required for HIF1A to transcriptionally activate the VEGFA gene and exert angiogenic activity; hypoxia increases ZFHX3 transcription via HIF1A binding to the ZFHX3 promoter (ChIP), and ZFHX3 loss blocks HIF1A-driven VEGFA expression, HUVEC migration/tube formation, and microvessel formation in HCC xenografts. |
Co-immunoprecipitation (ZFHX3-HIF1A), ChIP (HIF1A on ZFHX3 promoter), siRNA knockdown, luciferase reporter, HUVEC tube formation/migration assays, xenograft tumor assay |
The Journal of biological chemistry |
High |
32277050
|
| 2021 |
ZFHX3 transcriptionally represses SCN5A (encoding Nav1.5 sodium channel), reduces cardiac sodium current (INa) density in HL-1 cardiomyocytes, and increases expression of Nedd4-2 ubiquitin ligase to enhance Nav1.5 proteasomal degradation. ZFHX3 also reduces transcriptional activity of SCN5A, PITX2, TBX5, and NKX25 minimal promoters. |
Transfection/siRNA in HL-1 cells, patch clamp (INa measurement), luciferase promoter-reporter assay, Western blot |
International journal of molecular sciences |
Medium |
34884836
|
| 2021 |
Androgen/AR bidirectionally regulates ZFHX3 transcription in prostate cancer cells: androgen-induced AR binding to AREs in the ZFHX3 promoter upregulates ZFHX3 transcription, while ligand-free AR binds a distinct ARE to repress ZFHX3 transcription. Castration reduces Zfhx3 mRNA/protein in mouse prostates. |
ChIP (AR occupancy on ZFHX3 promoter in androgen-present vs. androgen-absent conditions), RT-PCR, Western blot, in vivo castration model |
Journal of cellular and molecular medicine |
Medium |
34953044
|
| 2023 |
Cardiomyocyte-restricted Zfhx3 loss causes atrial dilation, increased AF susceptibility (gene-dose dependent), altered conduction velocity, atrial action potential duration changes, calcium handling abnormalities, atrial thrombus, and dilated cardiomyopathy. SNP rs12931021 modulates an enhancer regulating ZFHX3 expression, with the AF risk allele associated with decreased ZFHX3 transcription (demonstrated by CRISPR editing and luciferase assay in iPSC-derived cardiomyocytes). |
CRISPR-Cas9 genome editing, chromatin immunoprecipitation, luciferase assay, conditional cardiac KO (hetero- and homozygous), echocardiography, MRI, electrophysiology, calcium imaging, RNA-seq, snATAC-seq analysis |
Circulation research |
High |
37449401
|
| 2024 |
A heterozygous GGC-repeat expansion in the last coding exon of ZFHX3 (encoding polyglycine) causes spinocerebellar ataxia type 4 (SCA4). Intranuclear ZFHX3-p62-ubiquitin aggregates are abundant in SCA4 basis pontis neurons. In patient fibroblasts and iPSCs, the GGC expansion leads to increased ZFHX3 protein levels and impairs autophagy; siRNA-mediated ZFHX3 knockdown normalizes both autophagy and protein levels. |
Long-read single-strand whole-genome sequencing, immunohistochemistry (intranuclear aggregates), autophagy assays in fibroblasts and iPSCs, siRNA rescue experiments |
Nature genetics |
High |
38684900
|
| 2024 |
Loss-of-function ZFHX3 variation causes syndromic intellectual disability; ZFHX3 interacts with the BRG1/Brm-associated factor (BAF) chromatin remodeling complex and the cleavage and polyadenylation complex (identified by co-immunoprecipitation/mass spectrometry confirmed by reverse co-IP and Western blot). ChIP-seq revealed ZFHX3 predominantly binds promoters of genes involved in nervous system development. |
Immunoprecipitation followed by mass spectrometry, reverse co-IP, Western blot, ChIP-seq, DNA methylation profiling, Drosophila knockdown model |
American journal of human genetics |
High |
38412861
|
| 2024 |
ZFHX3 acts as a tumor suppressor in prostate cancer by repressing FTO transcription; ZFHX3 knockdown increases FTO expression and thereby decreases global m6A RNA modification levels. FTO-mediated m6A demethylation regulates E2F2 and CDKN2C expression. Conversely, FTO regulates ZFHX3 expression in an m6A-dependent manner, establishing a bidirectional regulatory loop. |
siRNA knockdown, RT-PCR, Western blot, m6A quantification, MeRIP-seq, transcriptome sequencing, luciferase reporter assay (FTO promoter) |
Cell death discovery |
Medium |
38871709
|
| 2024 |
Myeloid-specific Zfhx3 deletion blocks hypercapnia-induced suppression of innate immune gene expression in alveolar macrophages and reduces influenza A virus replication, lung injury, and mortality in hypercapnic mice, establishing Zfhx3 as a mammalian mediator of CO2 effects on immune gene expression. |
Myeloid-specific conditional knockout (Zfhx3 flox × LysM-Cre), RNA-seq of alveolar macrophages, influenza A infection model with viral titer/lung injury measurement |
JCI insight |
High |
38227369
|
| 2025 |
ZFHX3 protein physically binds AR via multiple regions (particularly residues 1–223 of ZFHX3); ZFHX3 loss weakens AR transactivity and AR binding to target gene promoters (e.g., KLK3, FKBP5, TMPRSS2), and attenuates enzalutamide's anti-proliferative efficacy, establishing ZFHX3 as integral to AR signaling in prostate epithelial cells. |
Co-immunoprecipitation (ZFHX3-AR), ChIP (AR on target promoters), RNA-seq, luciferase reporter assay, cell proliferation/colony formation assay |
Scientific reports |
Medium |
40596315
|