| 1993 |
Mouse MTF-1 was cloned as a 72.5 kDa protein containing six zinc fingers and separate acidic, proline-rich transcriptional activation domains; ectopic expression strongly enhances transcription driven by MRE consensus sites or the MT-I promoter, and MTF-1–DNA complex formation is elevated several-fold in zinc-treated cells. |
cDNA cloning, bandshift (EMSA) assay, transfection reporter assay in primate/rodent cells |
The EMBO journal |
High |
8467794
|
| 1994 |
MTF-1 is required for metal-induced transcription via MREs; antisense MTF-1 prevents induction by all tested metals (zinc, cadmium, bismuth, silver, cobalt, copper, mercury, nickel), and ectopic MTF-1 causes constitutive MRE-reporter expression. Regulation is proposed to involve zinc releasing MTF-1 from an inhibitor. |
Stable transfection of MRE-reporter construct, antisense MTF-1 expression, cell fusion complementation assay |
Proceedings of the National Academy of Sciences of the United States of America |
High |
8108390
|
| 1994 |
Human MTF-1 was cloned as a 753 amino acid protein with 93% identity to mouse MTF-1, containing six zinc fingers; both species' proteins bind to MRE consensus sequences when expressed in mammalian cells, but human MTF-1 is more effective than mouse MTF-1 in mediating zinc-induced transcription. |
cDNA cloning, transfection reporter assay, chromosomal mapping |
Nucleic acids research |
High |
8065932
|
| 1995 |
Functional domain mapping of MTF-1 revealed three C-terminal transactivation domains (acidic, proline-rich, serine/threonine-rich) that function constitutively when fused to GAL4. The zinc finger region confers limited (3–4 fold) zinc-responsive transcription when fused to VP16, and the acidic/proline-rich activation domains also contribute to metal inducibility only in intact MTF-1 context. |
Deletion and chimeric mutant analysis, GAL4 fusion reporter assays, transfection in mammalian cells |
Nucleic acids research |
High |
7610056
|
| 1998 |
MTF-1 null mice die in utero at ~E14 with liver degeneration; MTF-1−/− embryos fail to transcribe metallothionein I/II genes and show diminished gamma-glutamylcysteine synthetase transcripts; primary MTF-1−/− fibroblasts show increased susceptibility to cadmium and hydrogen peroxide cytotoxicity. |
Targeted gene disruption (knockout mouse), RT-PCR/Northern blot of target genes, cytotoxicity assays in primary fibroblasts |
The EMBO journal |
High |
9582278
|
| 2000 |
MTF-1 is essential for both basal and metal (zinc and cadmium)-induced transcription of the ZnT1 (zinc transporter 1) gene; MTF-1 binds in vitro to the two MRE sequences in the ZnT1 promoter, demonstrated using MTF-1-null mouse embryo fibroblasts. |
In vitro DNA-binding assay (EMSA), MTF-1-null fibroblasts, Northern blot |
The Journal of biological chemistry |
High |
10952993
|
| 2001 |
Drosophila MTF-1 (dMTF-1) binds to conserved MREs and requires zinc for DNA binding, similar to mammalian MTF-1; dMTF-1 RNAi abolishes metal induction of both Drosophila metallothionein genes MtnA and MtnB, confirming its essential role in MT gene regulation. |
Transfection assays, EMSA, double-stranded RNA interference (RNAi) in Drosophila cells |
Molecular and cellular biology |
High |
11416130
|
| 2001 |
MTF-1 cooperates with USF1 for optimal MT-I gene expression in visceral endoderm cells; MTF-1 is absolutely required for upregulation of MT-I in visceral endoderm, and this expression depends on maternal dietary zinc as the signaling ligand. |
MTF-1 null mutant mouse embryos, USF1 null mutant analysis, zinc-deficient diet experiments |
The EMBO journal |
High |
11230134
|
| 2002 |
MTF-1 phosphorylation plays a critical role in its activation by zinc and cadmium; protein kinase C, tyrosine kinase, and casein kinase II are essential for metal-inducible transcriptional activation, while cAMP-dependent PKA is not directly involved. Phosphorylation does not regulate MTF-1 DNA binding; PKC inhibition results in elevated MTF-1 phosphorylation, suggesting specific dephosphorylation contributes to activation. |
Kinase inhibitor studies, transcriptional reporter assays, phosphorylation analysis |
The Journal of biological chemistry |
Medium |
11923282
|
| 2003 |
Drosophila MTF-1 knockout (by homologous recombination) abolishes basal and metal-induced expression of all four Drosophila metallothionein genes (MtnA, MtnB, MtnC, MtnD); MTF-1 null larvae are sensitive to both copper excess and copper starvation, with copper depletion blocking metamorphosis. |
Homologous recombination knockout in Drosophila, Northern blot/RT-PCR of MT genes, copper stress assays |
The EMBO journal |
High |
12505988
|
| 2004 |
MTF-1 is essential for embryonic liver formation (cell-autonomous defect demonstrated by co-cultivation failure) and for heavy metal detoxification in adult liver; conditional liver/bone marrow knockout mice are viable but highly susceptible to cadmium; MTF-1 deficiency in hematopoietic system causes severe lymphocyte underrepresentation. |
Conditional Cre-lox knockout (liver/bone marrow specific), co-cultivation assay of null vs. wild-type hepatocytes, cadmium challenge |
FASEB journal |
High |
15226267
|
| 2004 |
The individual zinc finger domains of MTF1 have differential metal ion affinities; the order from highest to lowest affinity is ZF4 > ZF2 ≈ ZF5 > ZF6 ≈ ZF3 ≈ ZF1, with domains at both termini showing lower intrinsic affinities than central domains; none of the six Cys2His2 zinc fingers has sufficiently low affinity to serve as a sensor of free micromolar zinc concentrations. |
In vitro cobalt(II)/zinc(II) metal ion affinity measurements with isolated single zinc finger peptides and His-to-Cys point mutants in full six-finger context |
Biochemistry |
High |
15122909
|
| 2004 |
Induction of metallothionein-I by phenolic antioxidants (tBHQ) requires MTF-1 and zinc; tBHQ activates MT1 through zinc-dependent MTF-1 binding to MREs, and tBHQ mobilizes intracellular zinc pools as demonstrated by fluorescence imaging; MTF-1-null cells are not rescued by tBHQ unless MTF-1 is re-expressed. |
MTF-1 null cell reconstitution, MRE-reporter assay, intracellular zinc fluorescence imaging, ZnT1-overexpressing cells |
The Biochemical journal |
High |
14998373
|
| 2005 |
In Drosophila, MTF-1 mediates transcriptional activation of Ctr1B (copper importer) upon copper depletion, in addition to activating metallothioneins upon copper excess; both responses depend on MRE motifs in the respective gene promoters, establishing MTF-1 as a bidirectional regulator of copper homeostasis. |
Reporter gene assay, MTF-1 knockout Drosophila, promoter MRE mutation analysis |
Genes & development |
High |
15833915
|
| 2006 |
The unique linker peptide RGEYT between zinc fingers 1 and 2 of MTF-1 is essential for zinc-sensing; replacing it with the canonical TGEKP linker results in constitutive DNA binding, nuclear translocation, and MT-I transcription regardless of zinc. The linker between ZF3 and ZF4 (TGKT) also contributes to zinc sensitivity. This establishes that zinc modulates linker-mediated zinc finger interactions to control MTF-1 activity. |
Linker swap mutagenesis, nuclear translocation assays, in vitro DNA binding, MT-I reporter gene assays in mammalian cells |
Molecular and cellular biology |
High |
16847313
|
| 2006 |
Nitric oxide donors (SNAP) cause nuclear translocation of MTF-1 in mouse lung endothelial cells; this NO-induced translocation requires metallothionein, as it does not occur in MT-knockout cells, indicating that NO releases zinc from MT zinc-thiolate clusters, raising labile zinc that activates MTF-1. |
EGFP-MTF-1 fusion nuclear translocation assay, MT-knockout endothelial cells, Western blot for MT protein |
Vascular pharmacology |
Medium |
16423564
|
| 2007 |
In zebrafish, zinc and cadmium mediate cytoplasmic-to-nuclear translocation of MTF-1–EGFP fusion protein in liver cells; recombinant zebrafish MTF-1 binds to MREs in the MT gene promoter in response to zinc and cadmium, as demonstrated by EMSA. |
EGFP-MTF-1 nuclear translocation fluorescence assay, recombinant protein EMSA, whole-mount in situ hybridization |
Environmental toxicology and chemistry |
Medium |
17269467
|
| 2008 |
Zinc treatment induces formation of a multiprotein coactivator complex containing MTF-1, histone acetyltransferase p300/CBP, and transcription factor Sp1; the acidic domain of MTF-1 is required to recruit p300 (but not Sp1); p300 knockdown reduces zinc-dependent MT-I transcription; the acidic domain also contains a nuclear exclusion signal (NES) with leucine residues critical for p300 recruitment and MT-I activation. |
Co-immunoprecipitation, siRNA knockdown of p300, acidic domain mutagenesis (L→A), NMR characterization of isolated acidic domain, reporter gene assays |
Molecular and cellular biology |
High |
18458062
|
| 2009 |
Within the MTF-1 DNA-binding domain, zinc fingers 1–3 (aa 137–228 in human MTF-1) harbor a nonconventional nuclear localization signal that confers constitutive nuclear localization on a cytoplasmic marker; the acidic activation domain contains a classical NES controlling cytoplasmic export; the acidic domain confers metal inducibility in distinct cell types when fused to a heterologous DNA-binding domain. A 3-aa difference in the NES explains the stronger metal inducibility of human vs. mouse MTF-1. |
Domain deletion/swap mutants fused to cytoplasmic marker, subcellular fractionation, reporter assays, chimeric human-mouse MTF-1 constructs |
Molecular and cellular biology |
High |
19797083
|
| 2010 |
Zinc and cadmium induce FPN1 (ferroportin) transcription through MTF-1; zinc causes MTF-1 nuclear translocation and binding to the FPN1 promoter; two functional MTF-1 binding sites in the mouse FPN1 promoter are required for zinc/cadmium-dependent expression; MTF-1 silencing reduces FPN1 transcription in response to zinc but not iron; Fpn1 protein can transport zinc and protects zinc-sensitive cells from zinc toxicity. |
MTF-1 nuclear translocation assay, siRNA silencing of MTF-1, promoter MRE site mutagenesis, reporter assay, zinc transport functional assay |
Blood |
High |
20688958
|
| 2010 |
MTF-1 mediates zinc-dependent regulation of ZnT2 expression in pancreatic acinar cells via an MRE downstream of the ZnT2 transcription start site; STAT5 and glucocorticoid receptor regulate ZnT2 via upstream STAT5 binding sites; ZnT2 knockdown increases cytoplasmic zinc and decreases zymogen granule zinc. |
Promoter deletion/mutation analysis, siRNA knockdown of ZnT2, fluorophore zinc measurement, reporter assays |
Proceedings of the National Academy of Sciences of the United States of America |
Medium |
20133611
|
| 2010 |
MTF-1 is essential for zinc-induced epigenetic changes at the MT-I promoter, including rapid decrease in Lys4-trimethylated and Lys9-acetylated histone H3 and loss of total histone H3 (nucleosome removal); an MTF-1 deletion mutant that binds promoter but cannot recruit p300 does not affect histone H3 in response to zinc, indicating nucleosome disruption requires an active MTF-1-coactivator complex. |
Chromatin immunoprecipitation (ChIP) for histone modifications, micrococcal nuclease sensitivity assay, MTF-1-null cells, p300-non-recruiting MTF-1 mutant |
Biochimica et biophysica acta |
High |
21035574
|
| 2011 |
Human MTF-1 homodimerizes via a conserved C-terminal cysteine cluster (632CQCQCAC638); dimer formation in vivo is important for basal and especially metal-induced transcriptional activity; copper stabilizes the dimer via intermolecular disulfide bonds and synergizes with zinc to boost MTF-1-dependent transcription; neither nuclear translocation nor DNA binding is impaired by cysteine-to-alanine mutation of this cluster. |
Co-immunoprecipitation (dimerization assay), cysteine cluster mutagenesis (C→A), reporter gene assays, copper treatment |
Biochimica et biophysica acta |
High |
22057392
|
| 2011 |
In Drosophila, MTF-1 partially rescues the parkin mutant phenotype (improved lifespan, motility, muscle/mitochondrial structure, female fertility); double mutant parkin/MTF-1 is synthetically lethal, placing MTF-1 in a complementary pathway to Parkin in metal homeostasis, oxidative stress, and cellular stress response. |
Drosophila genetic epistasis (double mutant), MTF-1 overexpression transgene, human MTF-1 transgene rescue, muscle/mitochondrial morphology assays |
Molecular and cellular biology |
High |
21383066
|
| 2011 |
MTF-1 activates Drosophila Ctr1B (copper importer) upon copper starvation through a C-terminal region including a cysteine-rich metallothionein-like domain; the C-terminus is autorepressive in the absence of copper; a phosphorylation site mutant (T127A) differentially affects target genes; constitutively active C-terminally truncated MTF-1 indiscriminately activates both metallothionein genes and Ctr1B. |
GAL4 fusion domain mapping, transgenic fly expression of truncated MTF-1 variants, phenotypic analysis |
The international journal of biochemistry & cell biology |
Medium |
22138226
|
| 2012 |
In Drosophila, MTF-1 selects different genomic DNA binding sites depending on the specific metal challenge (zinc vs. copper vs. cadmium), explaining metal-specific transcriptional responses; a single nucleotide in the MRE binding site can determine metal-specific transcription activation. |
Genome-wide MTF-1 ChIP mapping under different metal stresses, promoter reporter mutagenesis |
Proceedings of the National Academy of Sciences of the United States of America |
High |
23012419
|
| 2013 |
MTF-1 knockdown ameliorates zinc-induced lysosomal enlargement in TRPML1-deficient cells, placing MTF-1 downstream of TRPML1-mediated zinc release into the cytoplasm in a pathway involving ZnT4-mediated zinc compartmentalization. |
siRNA knockdown of MTF-1 and TRPML1, LysoTracker and zinc staining, lysosomal size quantification |
The Biochemical journal |
Medium |
23368743
|
| 2014 |
In osteoarthritis, ZIP8-mediated zinc influx activates MTF-1, which drives expression of matrix-degrading enzymes (MMP3, MMP9, MMP12, MMP13, ADAMTS5); genetic modulation of Mtf1 in mouse cartilage (intra-articular adenoviral expression or conditional knockout) alters OA pathogenesis, establishing the zinc-ZIP8-MTF1 axis as a catabolic regulator. |
Adenoviral overexpression and conditional knockout of Mtf1 in mouse cartilage, intra-articular injection model of OA, gene expression analysis |
Cell |
High |
24529376
|
| 2015 |
HIF-2α activates the zinc-ZIP8-MTF1 axis in chondrocytes by upregulating ZIP8, increasing zinc influx, and activating MTF1; reciprocally, the zinc-ZIP8-MTF1 axis transcriptionally regulates HIF-2α, creating a positive feedback loop that amplifies catabolic signaling in OA cartilage destruction. |
Conditional knockout mice for HIF-2α, ZIP8, and MTF1 in cartilage; adenoviral overexpression; microarray and gene expression assays |
Osteoarthritis and cartilage |
High |
26241779
|
| 2016 |
MTF1 controls a buffering hierarchy in zinc homeostasis: it activates metallothioneins and ZnT1 whose products buffer intracellular zinc, thereby dampening transcriptomic responses of other zinc-regulated genes; MTF1 knockdown increases the number and magnitude of zinc-responsive gene changes, while metallothionein overexpression curtails responses of zinc-repressed genes. |
siRNA knockdown of MTF1 in Caco-2 cells, transcriptome profiling, metallothionein overexpression, promoter reporter assay |
Metallomics |
Medium |
26824222
|
| 2018 |
MTF1 knockout in ovarian cancer cells (CRISPR/Cas9) inhibits epithelial-to-mesenchymal transition (EMT), upregulates E-cadherin and cytokeratin 7, downregulates Snai2 and β-catenin, reduces cell proliferation/migration/invasion, upregulates KLF4, and attenuates ERK1/2 and AKT survival signaling. |
CRISPR/Cas9 lentiviral knockout, EMT marker expression, migration/invasion assays |
Journal of Cancer |
Medium |
30588241
|
| 2018 |
A homozygous promoter variant in ATP7B that disrupts an MTF1 binding site causes reduced ATP7B expression in response to copper and likely causes Wilson Disease, demonstrating that MTF1 directly regulates ATP7B transcription in response to copper. |
Whole genome sequencing, bioinformatic MRE site analysis, in vitro reporter assay for promoter activity |
European journal of human genetics |
Medium |
30087448
|
| 2019 |
MTF1 binds specifically to MREe in the ATP7B promoter (validated by EMSA), and this interaction increases ATP7B promoter-driven reporter gene expression, establishing MTF1 as a direct transcriptional activator of ATP7B. |
Electrophoretic mobility shift assay (EMSA), MatInspector binding prediction, luciferase reporter assay |
Annals of human genetics |
Medium |
31596515
|
| 2019 |
MTF1 promotes myogenesis in response to copper; upon myogenesis initiation, MTF1 expression and nuclear localization increase; Mtf1 knockdown impairs differentiation; copper binds stoichiometrically to a C-terminal tetra-cysteine motif of MTF1; MTF1 binds chromatin at myogenic gene promoters and forms a complex with MYOD1 at these sites; copper supplementation stimulates MTF1 chromatin binding. |
shRNA and CRISPR/Cas9 knockdown, immunofluorescence, ChIP-seq, subcellular fractionation, atomic absorption spectroscopy, co-immunoprecipitation of MTF1-MYOD1 complex |
FASEB journal |
High |
31690123
|
| 2021 |
MTF-1 translocates to the nucleus in brain ischemia and directly binds the MRE at −23/−17 bp of the Ncx1 brain promoter, activating NCX1 transcription; MTF-1 silencing prevents RLIP-induced NCX1 upregulation and reduces neuroprotection (enlarged infarct volume), placing MTF-1 as an upstream activator of NCX1 in ischemic neuroprotection. |
MTF-1 siRNA silencing in rat ischemia model, chromatin immunoprecipitation (ChIP) for MTF-1 binding to Ncx1 promoter, infarct volume measurement |
Cell death & disease |
Medium |
33931586
|
| 2021 |
MTF1 activates Cacna1h transcription in hippocampal COCH neurons; elevated Cacna1h expression enables burst action potential firing in COCH neurons and mediates social-stress-induced anxiety-like behaviors via direct synaptic input to lateral septal GABAergic neurons. |
Genetic targeting of COCH neurons, MTF1 activity assays, electrophysiology, circuit tracing |
Cell reports |
Medium |
34965426
|
| 2022 |
The Hippo pathway kinases LATS1/2 phosphorylate and inhibit MTF1, reducing heavy metal response gene transcription and cellular protection; LATS activity is itself inhibited by accumulated zinc that directly binds and inhibits LATS; this reveals a zinc–LATS–MTF1 regulatory axis where zinc both activates MTF1 transcription (by relieving LATS inhibition) and inhibits LATS. |
Hippo pathway kinase overexpression/knockout, in vitro phosphorylation of MTF1 by LATS, heavy metal response gene reporter assays, zinc-LATS direct binding assay |
Nature cell biology |
High |
35027733
|
| 2022 |
PiC2 (SLC25A3), the mitochondrial Cu+ transporter, is a transcriptional target of MTF1; PiC2 expression is induced during myogenesis and by copper supplementation in an MTF1-dependent manner; PiC2 co-localizes and co-immunoprecipitates with COX, suggesting it participates upstream in copper delivery to cytochrome c oxidase. |
CRISPR/Cas9 knockout of PiC2, ChIP for MTF1 at PiC2 promoter, Co-IP of PiC2 and COX, in vitro Cu+-transfer assay |
Frontiers in molecular biosciences |
Medium |
36438658
|
| 2022 |
METTL3 acetylation (induced by sulfatide) reduces METTL3 binding to MTF1 mRNA, decreasing m6A methylation of MTF1 mRNA, prolonging its half-life and increasing MTF1 protein expression; mutation of 7 m6A methylation sites in the 3'UTR of MTF1 mRNA enhanced its stability, establishing m6A modification as a post-transcriptional regulatory mechanism controlling MTF1 expression. |
m6A quantification, METTL3 acetylation assay, MTF1 mRNA stability assay (half-life measurement), 3'UTR mutation reporter |
Chemistry & biodiversity |
Medium |
36149370
|