| 1990 |
LSF (TFCP2) was purified as a 63-kDa polypeptide that binds two distinct bipartite sites within the SV40 late promoter (LSF-GC site and LSF-280 site), recognizing directly repeated GC motifs with a center-to-center spacing of 10 bp, and activates transcription from the SV40 late promoter in vitro via binding to template DNA at initiation site L325. |
Protein purification, DNA affinity chromatography, methylation interference, in vitro transcription assay |
Genes & development |
High |
2159933
|
| 1992 |
CP2 (TFCP2) was molecularly cloned; bacterial expression of the full-length 502-aa murine factor or a core region (aa 133–395) generated polypeptides with the DNA-binding specificity of CP2, delimiting the region sufficient for specific DNA sequence recognition. The factor activates transcription of alpha-globin promoter templates ~3–4 fold in vitro. |
cDNA cloning, bacterial expression, DNase I footprinting, EMSA, in vitro transcription |
Molecular and cellular biology |
High |
1732747
|
| 1992 |
LSF (TFCP2) activates RNA polymerase II transcription by increasing the rate of association of TFIIB with the committed template on the SV40 major late promoter; LSF has no effect on TFIID binding rate or stability, but specifically accelerates the TFIIB assembly step. |
In vitro transcription kinetics, band mobility shift assays, general transcription factor titration |
The Journal of biological chemistry |
High |
1313810
|
| 1993 |
CP2 (TFCP2) binds a hyphenated recognition sequence motif spanning one DNA helix turn; elevated CP2 levels (2.5–5.5-fold) in stably transformed cells increased alpha-globin promoter-driven reporter activity 3–6 fold in vivo, confirming direct transcriptional activation. |
DNase I footprinting, EMSA, stable/transient transfection, CAT reporter assay |
The Journal of biological chemistry |
High |
8349681
|
| 1994 |
LSF (TFCP2) binds DNA as a homodimer, as demonstrated by an epitope-counting method. The alternatively spliced isoform LSF-ID lacks two beta-strand-forming regions conserved with Drosophila Elf-1/NTF-1; double amino acid substitutions in each of these regions eliminated specific DNA-binding activity. Mutations in one homology region, but not the other, eliminated dimerization. |
Epitope-counting EMSA, mutagenesis, protein-protein interaction studies |
Molecular and cellular biology |
High |
8035790
|
| 1994 |
The DNA-binding and dimerization domains of Drosophila Grainyhead are conserved in human CP2 (TFCP2). The DNA-binding domain (263 aa) contains a smaller core; the major dimerization domain is in a separate region and is required to stabilize DNA interactions. An N-terminal inhibitory domain can modulate activity. |
Domain deletion mapping, DNA-binding assays, dimerization assays in Drosophila and vertebrate (CP2) proteins |
Molecular and cellular biology |
High |
8196641
|
| 1995 |
CP2 (TFCP2) is pivotal to, but not sufficient for, the stage selector protein (SSP) complex that binds the stage selector element (SSE) in the gamma-globin promoter. CP2 requires a heterodimeric partner (~40–45 kDa) present in K562 cells to bind the SSE. The SSP complex is disrupted by anti-CP2 antiserum. The analogous chicken complex includes NF-E4, which is homologous to CP2. |
Biochemical purification, EMSA, UV and protein cross-linking, anti-CP2 antiserum disruption, Western blot |
The EMBO journal |
High |
7828600
|
| 1997 |
LSF (TFCP2) and YY1 cooperate to form a complex that recognizes the initiation region of the HIV-1 LTR and cooperatively repress LTR transcription and viral replication; transient expression of LSF alone has no effect, but cooperation with YY1 is required for repression in vivo. |
EMSA, transient transfection, viral replication assay |
Journal of virology |
High |
9371597
|
| 1997 |
LSF (TFCP2) DNA-binding activity is rapidly and quantitatively increased (~5-fold within 15 min) by mitogenic stimulation of resting human peripheral T lymphocytes, correlating with phosphorylation-induced reduction in LSF electrophoretic mobility. ERK1 (pp44) phosphorylates LSF in vitro at the same residue phosphorylated in vivo (position 291). Phosphatase treatment in vitro increases LSF mobility and decreases DNA-binding activity, confirming that phosphorylation directly increases DNA-binding. |
EMSA, in vitro kinase assay, mutagenesis (S291 mutant), phosphatase treatment, primary T lymphocyte stimulation |
Genes & development |
High |
9192871
|
| 1998 |
LSF (TFCP2) binds DNA as a tetramer (larger than a dimer) for high-affinity binding, requiring 234–320 amino acids. NTF-1 (Drosophila ortholog) can bind an LSF half-site as a dimer, whereas LSF cannot, indicating different oligomerization requirements despite shared DNA-binding domains. |
Truncation mutagenesis, EMSA, cross-linking experiments, mixing assays with full-length and truncation mutants |
The Journal of biological chemistry |
High |
9668115
|
| 1998 |
The Fe65 adaptor protein interacts with CP2/LSF/LBP1 (TFCP2) through its N-terminal PID1 domain, as shown by yeast two-hybrid and co-immunoprecipitation of native molecules. Different isoforms of CP2/LSF/LBP1 are distributed differently among subcellular fractions; at least one splice isoform (LSF-ID) is present outside the nucleus, while Fe65 is found in both nuclear and non-nuclear fractions. |
Interaction trap (yeast two-hybrid), co-immunoprecipitation, subcellular fractionation, transfection with HA-tagged constructs |
The Journal of biological chemistry |
High |
9685356
|
| 1998 |
Chicken CP2 (cCP2, TFCP2) forms a tetramer as shown by EMSA and cross-linking experiments. cCP2 is essential for transcription through the alphaA-crystallin lens enhancer alphaCE1; lens-specific enhancer activity depends on a 6-bp sequence (LSE) adjacent to the CP2 binding site, suggesting cooperation between cCP2 tetramer and a lens-specific factor binding LSE. |
EMSA, protein cross-linking, overexpression/dominant negative transfection, element substitution experiments |
Genes to cells |
High |
9753426
|
| 2000 |
LSF (TFCP2) binds sites within the thymidylate synthase (TS) promoter and intronic regions required for G1-S induction. Expression of dominant-negative LSF prevents the increase in TS enzyme levels during G1-S and induces S-phase-dependent apoptosis (thymineless death), which can be rescued by thymidine supplementation or heterologous TS expression. |
Reporter mutation analysis, dominant-negative expression, enzyme activity assay, apoptosis assays, rescue experiments |
The EMBO journal |
High |
10970859
|
| 2000 |
YY1 and LSF (TFCP2) cooperate to recruit HDAC1 to the HIV-1 LTR; HDAC1 co-purifies with the LTR-bound YY1-LSF repressor complex. LSF recruits YY1 to the LTR via YY1's zinc fingers (first two zinc fingers sufficient in vitro). The HDAC1-interacting domain of YY1 is required for LTR repression, and HDAC inhibitor trichostatin A blocks YY1-mediated repression. |
Co-purification, EMSA, HDAC inhibitor (TSA) treatment, deletion mutant analysis, HDAC1 overexpression |
Journal of virology |
High |
10888618
|
| 2000 |
Targeted disruption of CP2 (TFCP2) in mice produces viable animals with no apparent defects in hematopoiesis, globin gene expression, or immune responses. In CP2-null extracts, a slower-migrating complex bound to CP2 consensus sites is recognized by anti-NF2d9 (LBP-1a) antiserum, and recombinant LBP-1a can bind CP2 sites and form complexes with CP2 heteromeric partners, indicating functional compensation by the paralog LBP-1a/NF2d9. |
Gene targeting (knockout mice), EMSA with nuclear extracts, antiserum supershift, recombinant protein binding assay |
The Journal of biological chemistry |
High |
10995745
|
| 2001 |
Mutations in a putative CP2 (TFCP2) binding motif (-90C to A) in the uroporphyrinogen III synthase erythroid-specific promoter alter CP2 binding (EMSA) and reduce promoter activity to ~8% of wild-type in K562 cells, causing congenital erythropoietic porphyria. An adjacent -86A mutation does not alter CP2 binding. |
Sequencing, luciferase reporter assay, EMSA |
The Journal of clinical investigation |
High |
11254675
|
| 2003 |
LSF (TFCP2) is rapidly and quantitatively phosphorylated upon growth induction in NIH 3T3 cells; ERK is responsible for this phosphorylation, as shown by correlation of ERK activity with LSF phosphorylation and by specific MEK/ERK pathway inhibitors blocking LSF modification. ERK phosphorylation is necessary but not sufficient for activation of LSF DNA-binding activity (DNA-binding is not significantly altered in fibroblasts despite phosphorylation). |
Electrophoretic mobility shift, in vitro kinase assay, pharmacological inhibitors of Ras/Raf/MEK/ERK, cell fractionation |
Journal of cellular biochemistry |
High |
12858339
|
| 2005 |
CP2 (TFCP2) binds erythroid regulatory regions of the GATA-1, EKLF, and p45 NF-E2 genes and forms a ternary complex with GATA-1 and DNA at the GATA-1 HS2 enhancer. CP2 physically interacts with GATA-1 (demonstrated by co-immunoprecipitation and GST pull-down). Both factors occupy the same regulatory elements in vivo (chromatin IP). Mutation of the CP2 consensus in these elements severely impairs enhancer activity in K562 cells. |
EMSA, ChIP, co-immunoprecipitation, GST pull-down, transient transfection reporter assays |
Molecular and cellular biology |
High |
16648487
|
| 2005 |
Two HXPR-motif-containing proteins, REST and YY1, bind CP2 (TFCP2) via their HXPR sequences; GST pull-down and co-immunoprecipitation confirm this interaction. Double amino acid substitution in the HXPR sequence of YY1 abolishes binding to CP2 and eliminates YY1's suppression of CP2 transcriptional activity. At least three distinct binding regions on CP2 accommodate four different peptide motifs (HXPR, PHL, ASR, PXHXH). |
Peptide library screen, GST pull-down, co-immunoprecipitation, site-directed mutagenesis, transcriptional reporter assay |
The FEBS journal |
High |
15720400
|
| 2005 |
Erythroid cell-specific alpha-globin transactivation by CP2 (TFCP2) requires specific splice variants: CP2b (with an extra 36-aa exon) forms heteromeric complexes with CP2c; this heterodimer activates alpha-globin specifically in erythroid cells but not in non-erythroid 293T cells. Addition of PIAS1 (identified as a CP2-binding protein by yeast two-hybrid and confirmed by co-immunoprecipitation) restores alpha-globin activation in 293T cells, identifying PIAS1 as the missing erythroid cofactor. |
Yeast two-hybrid, co-immunoprecipitation, luciferase reporter in erythroid and non-erythroid cells, expression of recombinant isoforms |
Molecular and cellular biology |
High |
15988015
|
| 2006 |
APLP2 intracellular C-terminal domain (ICD) interacts with CP2 (TFCP2) in the nucleus and induces expression of GSK-3β. Nuclear colocalization and interaction were demonstrated, and GSK-3β protein levels increased upon APLP2-ICD expression. |
Co-immunoprecipitation, transfection, immunofluorescence, Western blot |
Cell death and differentiation |
Medium |
16645641
|
| 2009 |
LSF (TFCP2) is phosphorylated by cyclin C/CDK2 (and cyclin C/CDK3 in humans) predominantly at S309 during early G1 after mitogenic stimulation of quiescent fibroblasts. This phosphorylation is mediated by cyclin C-dependent kinases (confirmed by co-immunoprecipitation and siRNA knockdown of cyclin C), and phosphorylation at S309 inhibits LSF transactivation and prevents induction of the LSF target gene Tyms at G1/S. |
In vitro kinase assays, mutagenesis (S309), co-immunoprecipitation, siRNA knockdown of cyclin C, transient transfection reporter, endogenous Tyms expression |
Molecular and cellular biology |
High |
19237534
|
| 2005 |
LSF (TFCP2) binds the HIV-1 LTR as a tetramer; ERK phosphorylation of LSF decreases its binding to the LTR in vitro, while p38 phosphorylation increases binding. In cells, p38 agonist increases LSF occupancy at LTR chromatin; p38 inhibition decreases LSF occupancy, increases histone H4 acetylation at the adjacent nucleosome, blocks YY1-mediated LTR repression, and results in recovery of HIV from resting CD4+ T cells of aviremic donors. |
EMSA, ChIP, kinase assays, pharmacological p38 inhibition/agonism, HIV recovery assay from primary cells |
Journal of virology |
High |
15857981
|
| 2010 |
LSF (TFCP2) functions as an oncogene in hepatocellular carcinoma: LSF overexpression promotes aggressive, angiogenic, and multiorgan metastatic tumors in nude mice, while inhibition abrogates growth and metastasis. LSF transcriptionally upregulates osteopontin (OPN), which mediates key oncogenic functions; loss of OPN impairs LSF-driven tumorigenesis. |
Stable overexpression/dominant-negative, nude mouse xenograft, microarray, loss-of-function studies with siRNA/shRNA |
Proceedings of the National Academy of Sciences of the United States of America |
High |
20404171
|
| 2011 |
LSF (TFCP2) transcriptionally upregulates MMP-9 to enhance angiogenesis in HCC. ChIP-on-chip identified MMP-9 as a direct LSF target; ChIP confirmed LSF binding to the MMP-9 promoter. Deletion mutation analysis identified LSF-responsive promoter regions. MMP-9 inhibition abrogated LSF-induced angiogenesis and tumorigenesis in vivo. |
ChIP-on-chip, ChIP-PCR, promoter deletion reporter assays, HUVEC differentiation assay, CAM assay, nude mouse xenograft |
The Journal of biological chemistry |
High |
22167195
|
| 2011 |
LSF (TFCP2) activates c-Met signaling in HCC through a pathway involving LSF-driven OPN secretion activating c-Met via CD44. Co-immunoprecipitation confirmed OPN-induced CD44/c-Met interaction. Chemical or genetic inhibition of c-Met abrogates LSF-mediated tumorigenesis and metastasis in vivo. |
Phospho-RTK array, co-immunoprecipitation, tissue microarray IHC, siRNA, chemical inhibitor, nude mouse xenograft |
Journal of hepatology |
Medium |
21703197
|
| 2015 |
TFCP2 directly regulates FN1 (fibronectin 1) and TJP1 (tight junction protein 1) as transcriptional targets mediating HCC metastasis. ChIP-on-chip combined with transcriptome analysis identified these targets; ChIP-PCR confirmed TFCP2 binding to the FN1 promoter, and promoter reporter identified TFCP2-binding motifs. FN1 inhibition blocks TFCP2-induced aggression; TFCP2 overexpression rescues FN1 inhibition. |
ChIP-on-chip, transcriptome microarray, ChIP-PCR, promoter reporter assay, siRNA rescue experiments |
Journal of experimental & clinical cancer research |
High |
25609232
|
| 2016 |
LSF (TFCP2) inhibits melanoma cell growth by binding within a 150-bp upstream region of the p21CIP1 transcription start site and increasing p21CIP1 expression, resulting in G1 phase arrest. EMSA and ChIP assays confirmed LSF binding to the p21CIP1 promoter. |
EMSA, ChIP, overexpression/depletion, cell cycle analysis, anchorage-dependent/independent growth assays, xenograft |
Oncotarget |
High |
26506241
|
| 2017 |
TFCP2 is required for YAP-dependent transcription in liver cancer. Mechanistically, TFCP2 stimulates YAP function via a WW-PSY interaction, maintains YAP stability by inhibiting βTrCP-mediated ubiquitination, and acts as a transcription co-factor facilitating YAP binding to YBF-containing transcription factors and the YAP-TEAD interaction. Genomic co-occupancy of YAP and TFCP2 was revealed by ChIP. |
Co-immunoprecipitation (WW-PSY interaction), ubiquitination assays, ChIP, luciferase reporter, siRNA/overexpression |
Cell reports |
High |
29091762
|
| 2018 |
Crystal structures of the DNA-binding domains of Grainyhead-like (Grhl) 1 and Grhl2 (close CP2/TFCP2 family members) reveal an immunoglobulin-like core fold shared with p53, with dimeric DNA binding mediated by two conserved arginine residues. A cancer-associated mutation in Grhl1 at one of these arginines completely abrogates DNA binding in biochemical assays and transcriptional activation in human cells. |
X-ray crystallography, in vitro DNA-binding assays, cancer-mutation analysis, transcriptional reporter assay in human cell line |
Nucleic acids research |
High |
29309642
|
| 2017 |
TFCP2 promotes pancreatic cancer progression by interacting with β-catenin, enhancing the β-catenin/TCF4 interaction, and activating β-catenin/TCF signaling, as demonstrated by co-immunoprecipitation and luciferase reporter assay. |
Co-immunoprecipitation, luciferase reporter (β-catenin/TCF), overexpression/knockdown, colony formation, in vivo metastasis model |
Oncotarget |
Medium |
29050300
|
| 2019 |
CCT3 co-interacts with YAP and TFCP2 (confirmed by mass spectrometry co-immunoprecipitation) and prolongs their half-lives by blocking PCBP2-induced ubiquitination in a βTrCP-independent manner. PCBP2 directly interacts with YAP via a WB motif-WW domain interaction and indirectly with TFCP2 via YAP. CCT3 separates PCBP2-YAP interactions, preventing ubiquitination of both YAP and TFCP2. |
Mass spectrometry co-immunoprecipitation, ubiquitination assays, co-immunoprecipitation, Western blot half-life assay |
Cell death & disease |
Medium |
31501420
|
| 2020 |
TFCP2 directly bound the promoters of EGF and TGFα to regulate their expression, stimulating autocrine EGFR signaling and AKT activation in triple-negative breast cancer. This creates a positive feedback loop controlling TFCP2-mediated EMT, stemness, and metastasis. |
ChIP, promoter reporter assay, siRNA/overexpression, EGFR signaling assays, in vivo metastasis model |
Cancer research |
Medium |
32193292
|
| 2020 |
LSF (TFCP2) binds α-tubulin and SET8 (KMT5A/PR-Set7), facilitating SET8-mediated methylation of α-tubulin at Lys311. LSF enhances SET8-mediated α-tubulin methylation in vitro; the LSF inhibitor FQI1 counters this facilitation. SET8 bound α-tubulin and methylated it at Lys311 as shown by in vitro radiolabeling, MS, and immunoblotting. |
In vitro radiolabeling, mass spectrometry, immunoblotting, co-immunoprecipitation, immunofluorescence, GST pull-down, FQI1 inhibitor |
The Journal of biological chemistry |
High |
32111740
|
| 2020 |
Inhibition of LSF (TFCP2) by either the small molecule FQI1 or siRNA causes highly similar mitotic delays with condensed, unaligned chromosomes preceding metaphase, leading to multi-nucleation, apoptosis, and senescence. The concordance between FQI1 and siRNA phenotypes confirms LSF as the specific cellular target of FQI1. |
Time-lapse microscopy, bulk cell synchronization, siRNA knockdown vs. FQI1 treatment comparison, cell death/senescence assays |
BMC cancer |
High |
32539694
|
| 2009 |
TFCP2 acts as a transcription factor that directly binds the SRY promoter (demonstrated by EMSA); TFCP2 suppression by RNAi significantly reduced SRY mRNA expression in cells, while overexpression did not significantly increase SRY mRNA, indicating TFCP2 is required but not limiting for SRY expression. |
EMSA, RNAi knockdown, overexpression, RT-PCR |
Molecular and cellular biochemistry |
Medium |
19902333
|
| 2021 |
TFCP2 interacts with SREBP2 (co-immunoprecipitation) and synergistically activates expression of HMGCR (a rate-limiting enzyme in cholesterol synthesis), enabling pancreatic cancer cells to overcome oncogene-induced senescence. Statins (which inhibit HMGCR) reversed the anti-senescence effect of TFCP2. |
Co-immunoprecipitation, qRT-PCR, luciferase reporter, statin treatment, senescence assays (SA-β-gal) |
Frontiers in oncology |
Medium |
34804919
|
| 2022 |
TFCP2 induces SMAD2 expression as a transcription factor; ITGA2 inhibits the nuclear translocation of TFCP2, thereby reducing SMAD2 transcription and suppressing TGF-β pathway activation. Co-immunoprecipitation and nuclear/cytoplasmic fractionation confirmed that ITGA2 interacts with and retains TFCP2 in the cytoplasm. |
Co-immunoprecipitation, nuclear/cytoplasmic fractionation, ChIP, luciferase reporter, Western blot, RT-qPCR |
Journal of experimental & clinical cancer research |
Medium |
35193647
|
| 2022 |
TFCP2 interacts with ATF3 to cooperatively regulate de novo serine synthesis by promoting ATF3 binding to the PHGDH promoter. Knockdown of TFCP2 significantly inhibited ATF3 binding to the PHGDH promoter, as shown by ChIP. |
Co-immunoprecipitation, ChIP, siRNA knockdown, cell proliferation and sphere formation assays |
Experimental cell research |
Medium |
35421367
|
| 2024 |
TRAF6 stabilizes YAP1 by K63 poly-ubiquitination, promoting formation of the YAP1/TFCP2 transcriptional complex and subsequent PD-L1 transcription in melanoma. Suppression of TRAF6 down-regulates PD-L1 on the membrane surface of melanoma cells. In vitro and in vivo assays confirmed the TRAF6→YAP1 K63-ubiquitination→YAP1/TFCP2 complex→PD-L1 transcription axis. |
CRISPR interference screen, ubiquitination assays (K63-specific), co-immunoprecipitation, PD-L1 flow cytometry, luciferase reporter, in vitro/in vivo tumor models |
Cancer letters |
Medium |
38583649
|
| 2024 |
FUS-TFCP2 fusion protein blocks myogenic differentiation, induces transcription of ALK and truncated TERT, and inhibits DNA repair in rhabdomyosarcoma. Functional studies in cell models showed genomic instability and signs of defective homologous recombination in TFCP2-rearranged tumors. |
Functional cell-based studies with FUS-TFCP2 expression constructs, transcriptome analysis, DNA repair assays |
Nature communications |
Medium |
38168093
|
| 2017 |
CP2 (TFCP2) upregulates miR-144 expression in mouse ovarian granulosa cells by directly binding the miR-144 promoter, as shown by luciferase reporter, ChIP, and EMSA. CP2-driven miR-144 expression suppresses COX-2 and reduces PGE2 production. |
Luciferase reporter, ChIP, EMSA, miR-144 inhibitor/mimic overexpression |
Cell death & disease |
Medium |
28182010
|
| 2012 |
The small molecule FQI1 inhibits LSF (TFCP2) DNA-binding activity in vitro (EMSA) and in cells (ChIP), eliminates transcriptional stimulation of LSF-dependent reporter constructs, and induces rapid cell death in LSF-overexpressing HCC cells without affecting normal hepatocytes. Structure-activity relationships of 23 quinolinones are highly concordant between antiproliferative and LSF-inhibitory activities, confirming LSF as the specific target. |
EMSA, ChIP, luciferase reporter, cell viability assays, SAR analysis, xenograft mouse model |
Proceedings of the National Academy of Sciences of the United States of America |
High |
22396589
|