| 2008 |
CXXC5 co-localizes with Dishevelled (Dvl) in the cytoplasm and physically interacts with Dvl, as demonstrated by co-immunoprecipitation and FRET experiments. Overexpression of CXXC5 represses the canonical Wnt signaling target Axin2 and attenuates Wnt3a-mediated TOPflash reporter activity; RNAi knockdown of CXXC5 attenuates BMP4-mediated decrease in Axin2 levels, establishing CXXC5 as a BMP4-induced inhibitor of Wnt signaling in neural stem cells. |
Co-immunoprecipitation, FRET, TOPflash reporter assay, RNAi knockdown, Axin2 expression analysis |
The Journal of biological chemistry |
High |
19001364
|
| 2009 |
CXXC5 (RINF) encodes a nuclear factor containing a CXXC-type zinc-finger motif that is induced by retinoids in acute promyelocytic leukemia cells; shRNA knockdown demonstrates a regulatory function in normal and tumoral myelopoiesis. |
Microarray expression profiling, shRNA knockdown, nuclear localization by cell fractionation |
Blood |
Medium |
19182210
|
| 2013 |
CXXC5 is a transcriptional activator of the Flk-1 (VEGFR2) gene; in vitro DNA binding assay showed direct binding of CXXC5 to the Flk-1 promoter region, and mutation of the DNA-binding motif abolished transcriptional activity. BMP4 induces CXXC5 transcription which in turn induces Flk-1; CXXC5 knockdown suppressed BMP4-induced stress fiber formation and migration in HUVECs, and cxxc5 morpholino injection in zebrafish caused caudal vein plexus defects. |
In vitro DNA binding assay, promoter-reporter assay, site-directed mutagenesis, siRNA knockdown, morpholino zebrafish model, CXXC5−/− mouse Matrigel angiogenesis assay |
FASEB journal |
High |
24136587
|
| 2013 |
CXXC5 interacts with Smad proteins (Smad3 and Smad4) as shown by co-localization and co-immunoprecipitation; CXXC5 facilitates Smad3 phosphorylation and Smad4 nuclear translocation, and co-expression of Smad with CXXC5 increases TNF-α reporter activity, linking CXXC5 to the extrinsic apoptosis pathway. |
FRET, co-immunoprecipitation, nuclear fractionation, caspase activity assay, TUNEL assay, transcription reporter assay, Western blot |
Current molecular medicine |
Medium |
23906331
|
| 2015 |
CXXC5 negatively regulates osteoblast differentiation and bone formation via a Wnt-dependent interaction with Dvl. CXXC5-deficient mice exhibit elevated bone mineral density; a competitor peptide (PTD-DBM) blocking the CXXC5-Dvl interaction activates the Wnt/β-catenin pathway and accelerates ex vivo calvarial bone growth. |
CXXC5−/− mouse bone phenotyping (DEXA, micro-CT), co-immunoprecipitation of CXXC5-Dvl, competitor peptide functional assay, ex vivo calvarial culture |
Cell death and differentiation |
High |
25633194
|
| 2015 |
CXXC5 acts as a negative feedback regulator of Wnt/β-catenin signaling in skin by interacting with Dvl. CXXC5−/− mice show accelerated cutaneous wound healing with enhanced β-catenin, collagen I, and keratin 14. PTD-DBM competitor peptide disrupts CXXC5-Dvl interaction; co-treatment with PTD-DBM and valproic acid synergistically accelerates wound healing. |
CXXC5−/− mouse wound healing model, overexpression/knockdown of CXXC5 in vitro, PTD-DBM competitor peptide, β-catenin/collagen measurements |
The Journal of experimental medicine |
High |
26056233
|
| 2015 |
CXXC5 functions as a transcriptional activator of major myelin genes (including MBP) by directly binding the MBP promoter through its CXXC DNA-binding motif. CXXC5−/− mice show severely reduced myelin gene expression in corpus callosum, abnormal myelin structure, and reduced electrical conduction amplitudes. |
In vitro DNA binding assay (CXXC5 on MBP promoter), CXXC5−/− mouse myelin phenotyping (qPCR, electron microscopy, electrophysiology), neural stem cell differentiation assays |
Glia |
High |
26462610
|
| 2016 |
Crystal structure of the mouse Dvl1 PDZ domain (1.76 Å resolution) was determined, and molecular modeling using NMR and X-ray data defined the Dvl1 PDZ domain binding pocket for the CXXC5 peptide, providing structural basis for the CXXC5-Dvl interaction. |
X-ray crystallography (1.76 Å resolution), NMR, molecular modeling |
Biochemical and biophysical research communications |
High |
27932247
|
| 2016 |
Small-molecule inhibitors of the Dvl-CXXC5 interaction were identified and shown to activate the Wnt/β-catenin pathway and enhance osteoblast differentiation. NMR titration confirmed direct binding of compound KY-02061 to the Dvl PDZ domain. Oral administration of KY-02327 rescued bone loss in ovariectomized mice. |
Fluorescence polarization in vitro assay (Dvl-CXXC5 disruption), NMR titration, primary osteoblast differentiation assay, ex vivo calvaria culture, OVX mouse model |
EMBO molecular medicine |
High |
26941261
|
| 2016 |
CXXC5 expression is induced by Wnt/β-catenin signaling and in turn directly binds the MBP promoter to activate myelin gene expression, positioning CXXC5 as part of a Wnt-driven transcriptional program in oligodendrocytes. |
Luciferase reporter assay, ChIP/DNA binding assay on MBP promoter, Wnt pathway modulation in neural stem cells |
Glia |
Medium |
26462610
|
| 2016 |
CXXC5 induces H3K9 methylation at the Cd40lg promoter through association with the histone methyltransferase SUV39H1, thereby repressing CD40L expression in CD8+ cytotoxic T cells. ThPOK represses CXXC5 expression, thereby de-repressing CD40L. |
Retroviral Thpok transduction, CXXC5 transgene overexpression in T cells, chromatin modification analysis (H3K9me, H3K27me), co-immunoprecipitation of CXXC5 with SUV39H1 |
Journal of leukocyte biology |
Medium |
26896487
|
| 2016 |
CXXC5 expression is regulated by E2-ERα through a direct estrogen response element (ERE) upstream of the CXXC5 translation start codon, establishing CXXC5 as an E2-ERα responsive gene. |
ERE-luciferase reporter assay, ERα ChIP, site-directed mutagenesis of ERE, qPCR/Western blot with E2 treatment |
Scientific reports |
Medium |
27886276
|
| 2017 |
CXXC5 recruits DNA demethylase TET2 to maintain hypomethylation of CpG islands within IRF7 and other interferon pathway genes in plasmacytoid dendritic cells (pDCs). Genetic ablation of CXXC5 causes aberrant CpG methylation of the IRF7 gene locus, impairs IRF7 expression, and compromises TLR7/9- and virus-induced IFN response. This positions CXXC5 as an epigenetic regulator that anchors TET2 at specific CGIs. |
CXXC5 knockout mouse, bisulfite sequencing/methylation analysis of Irf7 CGI, TET2 recruitment assay (ChIP), IFN response assays, viral challenge model |
The Journal of experimental medicine |
High |
28416650
|
| 2017 |
CXXC5 is a negative regulator of the Wnt/β-catenin pathway in hair follicles via interaction with Dvl. CXXC5 is upregulated in miniaturized follicles in human balding scalps. CXXC5 inhibits alkaline phosphatase activity and cell proliferation in human hair follicle dermal papilla cells; CXXC5−/− mice show accelerated hair regrowth; disrupting the CXXC5-Dvl interaction with PTD-DBM peptide activates Wnt/β-catenin and accelerates hair regrowth and wound-induced hair neogenesis. |
CXXC5−/− mouse hair regrowth model, wound-induced hair neogenesis model, PTD-DBM competitor peptide treatment, alkaline phosphatase activity assay, cell proliferation assay in dermal papilla cells |
The Journal of investigative dermatology |
High |
28595998
|
| 2018 |
CXXC5 is a novel TGF-β target gene; knockdown of CXXC5 attenuates expression of a substantial portion of TGF-β target genes and ameliorates TGF-β-induced growth inhibition and apoptosis. CXXC5 associates with HDAC1 and competes with HDAC1 for interaction with Smad2/3, thereby abolishing the inhibitory effect of HDAC1 on TGF-β signaling. |
RNA-Seq identification of TGF-β target genes, CXXC5 knockdown, co-immunoprecipitation of CXXC5 with HDAC1 and Smad2/3, competition assay, cell growth/apoptosis assays |
Journal of molecular cell biology |
High |
29036306
|
| 2019 |
Rinf (CXXC5) binds to the chromatin at promoters and enhancers of Tet1, Tet2, and pluripotency genes in mouse ESCs, where it forms a complex with Nanog, Oct4, Tet1, and Tet2, and facilitates their recruitment to regulatory regions. Rinf deficiency reduces expression of pluripotency factors and Tet enzymes and causes aberrant differentiation. |
ChIP-seq (Rinf occupancy), Co-immunoprecipitation (Rinf with Nanog, Oct4, Tet1, Tet2), Rinf KO mouse ESCs, RNA-seq, differentiation assays |
Cell reports |
High |
31433977
|
| 2019 |
CXXC5 (RINF) mediates growth plate senescence via suppression of Wnt/β-catenin signaling through its interaction with Dvl. Cxxc5−/− mice show delayed growth plate senescence and tibial elongation. Indirubin analog KY19382, identified by an in vitro CXXC5-DVL interaction screening assay, disrupts this interaction and elongates tibial length. |
Cxxc5−/− mouse growth plate analysis, in vitro CXXC5-DVL interaction screening assay, tibial length measurement, growth plate histology |
Life science alliance |
Medium |
30971423
|
| 2019 |
CXXC5 plays a zebrafish cardiac looping role via TGF-β signaling; zebrafish CXXC5 interacts with SMAD through its ZF-CXXC domain and SMAD MH1 domain. Overexpression of CXXC5 increases TGF-β luciferase reporter activity; cxxc5 morpholino knockdown causes cardiac looping defects rescued by hand2 mRNA co-injection. |
Co-immunoprecipitation (CXXC5-SMAD domain interaction), TGF-β luciferase reporter assay, zebrafish morpholino knockdown, mRNA rescue experiment |
International journal of cardiology |
Medium |
27077543
|
| 2020 |
CXXC5 was verified by recombinant protein generation to directly bind unmethylated CpG dinucleotides. Despite this DNA-binding capacity, CXXC5 lacks intrinsic transcription activation/repression function but participates in E2-driven cellular proliferation by modulating expression of distinct and mutual genes regulated by E2. |
Recombinant CXXC5 protein production, in vitro unmethylated CpG binding assay, transcription activation/repression reporter assays (negative for intrinsic activity), gene expression profiling with CXXC5 overexpression/knockdown |
Scientific reports |
Medium |
32249801
|
| 2020 |
Cxxc5 expression affects cell cycle and myeloid differentiation of mouse HSCs: knockdown reduces monocyte and increases granulocyte development; Cxxc5 knockdown increases S-phase fraction and proliferation, whereas overexpression decreases S-phase fraction; RNA-seq identified upregulation of cell cycle regulators after knockdown. |
shRNA knockdown and lentiviral overexpression in mouse LSK cells, flow cytometry cell cycle analysis, ex vivo myeloid differentiation, RNA-seq, single-cell RNA-seq |
Journal of leukocyte biology |
Medium |
32083332
|
| 2021 |
CXXC5 binds the proximal MYCL1 promoter and represses MYCL1 transcription in quiescent hepatic stellate cells (HSCs). Loss of CXXC5 during HSC activation removes CpG methylation and acquires acetylated H3K9/H3K27 at the MYCL1 promoter, leading to MYCL1 transactivation and HSC activation. MYCL1 knockdown attenuates HSC activation; MYCL1 overexpression partially relieves CXXC5-mediated blockade. |
ChIP (CXXC5 on MYCL1 promoter), bisulfite sequencing, histone modification analysis, CXXC5 overexpression/knockout, RNA-seq, MYCL1 knockdown/overexpression functional assays |
Frontiers in cell and developmental biology |
High |
34621736
|
| 2021 |
Proximity-dependent biotinylation (BioID) mapping identified CXXC5 interaction partners including DNA/chromatin modifiers, transcription factors/co-regulators, and RNA processors. CXXC5 interacts through its CXXC domain with EMD, MAZ, and MeCP2; interplay between CXXC5 and MeCP2 was critical for a subset of CXXC5 target gene expressions. |
BioID proximity biotinylation, sequential immunoprecipitation coupled to mass spectrometry, domain-interaction mapping, gene expression analysis of CXXC5/MeCP2 co-regulated genes |
Scientific reports |
Medium |
34475492
|
| 2022 |
RINF (CXXC5) upregulates SMAD7 expression by direct binding to the SMAD7 promoter in immature erythroid cells, maintaining SMAD7 levels to fine-tune TGF-β sensitivity. RINF knockdown accelerates erythropoietin-driven maturation and reduces RBC numbers (~45%); ectopic SMAD7 expression rescues the RINF knockdown phenotype. RINF silencing also affects 5'-hydroxymethylation of erythroblasts, consistent with a Tet2-anchoring role. |
RINF shRNA knockdown in primary human CD34+ cells, promoter binding (ChIP), ectopic SMAD7 rescue experiment, 5-hydroxymethylcytosine profiling, erythroid differentiation assays |
Haematologica |
High |
33241676
|
| 2022 |
Idax and Rinf (CXXC5) facilitate expression of Tet enzymes to promote neural and suppress trophectoderm programs during ESC differentiation. Individual or combined loss of Idax and Rinf in ESCs downregulates Tet genes (direct targets) and neural markers, and upregulates trophectoderm markers. DKO embryos have reduced NPC markers in forebrain and deregulated trophoblast markers in placenta. |
Single and double knockout ESCs, directed differentiation to NPCs and trophoblast-like cells, DKO mouse embryo analysis (NPC/trophoblast markers), qPCR/Western blot |
Stem cell research |
Medium |
35390758
|
| 2022 |
CXXC5 interacts with the CRL4B and NuRD complexes and the CXXC5-CRL4B-NuRD complex mediates transcriptional repression of TSC1 and other genes, activating mTOR signaling and PD-L1 expression in breast cancer. ChIP-seq defined direct transcriptional targets of this complex. |
Co-immunoprecipitation (CXXC5 with CRL4B and NuRD components), ChIP-seq, loss-of-function (knockdown) and overexpression, in vitro proliferation and in vivo xenograft assays |
The Journal of biological chemistry |
Medium |
36539038
|
| 2022 |
BMP signaling promotes exit of DIPG tumor cells from a stem-cell-like state to differentiation by epigenetically regulating CXXC5, which acts as a tumor suppressor and positive regulator of BMP signaling in H3.3K27M/ACVR1-WT DIPG. |
BMP ligand treatment, CXXC5 gain/loss-of-function in DIPG cells, epigenetic profiling, SMAD-dependent reporter assays, in vivo DIPG models |
Nature cancer |
Medium |
35915262
|
| 2024 |
CXXC5 extensively co-localizes with TET1 and TET2 at CpG islands in mouse ES cells and interacts with 5-methylcytosine oxidases (TET proteins). CXXC5 knockout leads to substantial genome-wide DNA hypomethylation affecting all genomic compartments, consistent with a model in which CXXC5 anchors TET proteins at CpG islands and in its absence TET enzymes induce genome-scale demethylation. |
CXXC5 knockout (gene editing), whole-genome bisulfite sequencing, co-localization analysis of CXXC5 with TET1/TET2 (ChIP), RNA-seq |
Epigenomics |
High |
39585161
|
| 2024 |
CXXC5 drives ovarian cancer cell proliferation via transcriptional activation of ZNF143 and EGR1 downstream transcription factors. CXXC5 expression is directly regulated by HIF1A (hypoxia). Loss of CXXC5 inactivates multiple inflammatory signaling pathways; in vitro and in vivo experiments confirmed ZNF143 and EGR1 as downstream transcriptional targets mediating CXXC5's proliferative effects. |
CXXC5 knockdown/overexpression, HIF1A ChIP (CXXC5 promoter), downstream target gene expression (ZNF143, EGR1), in vitro proliferation assays, in vivo xenograft |
Cellular signalling |
Medium |
38642782
|
| 2025 |
CXXC5 protein is regulated post-translationally by ubiquitination and degraded via the ubiquitin-proteasome pathway. Specific ubiquitinated lysine residues of CXXC5 were identified by bioUbiquitination approach followed by sequential immunoprecipitation coupled to mass spectrometry; these lysine residues contribute to CXXC5 degradation in MCF-7 and HEK293FT cells. E2 augments CXXC5 transcription and synthesis in the G1 phase, but protein levels are primarily controlled by ubiquitination independently of cell cycle phase. |
BioUbiquitination approach (cellular biotinylation of ubiquitin), sequential immunoprecipitation coupled mass spectrometry, cell synchronization, cycloheximide chase, proteasome inhibitor treatment, Western blot |
Protein science |
High |
40371716
|