| 2018 |
CXorf67 (EZHIP) physically interacts with EZH2, SUZ12, and EED — core components of PRC2 — as detected by immunoprecipitation/mass spectrometry in the Daoy cell line. Enforced reduction of CXorf67 in Daoy cells restored H3K27me3 levels, while enforced expression in HEK293T and neural stem cells reduced H3K27me3 levels, establishing CXorf67 as a functional inhibitor of PRC2-mediated H3K27 trimethylation. |
Immunoprecipitation/mass spectrometry; gain- and loss-of-function experiments (knockdown and overexpression) with western blot readout of H3K27me3 |
Acta neuropathologica |
High |
29909548
|
| 2019 |
A conserved short peptide sequence in the C-terminal region of EZHIP (CXorf67) that mimics the H3 K27M oncohistone sequence is necessary and sufficient to bind the SET domain of EZH2 and inhibit PRC2 catalytic methyltransferase activity in vitro and in vivo, causing global loss of H3K27me2/3 and de-repression of PRC2 target genes including neurodevelopmental genes. |
Mass spectrometry; peptide modeling; immunocytochemistry; western blot; in vitro methyltransferase assay; domain mapping |
Neuro-oncology |
High |
30923826
|
| 2019 |
A conserved sequence in EZHIP directly contacts the active site of EZH2 in a mechanism analogous to H3 K27M. Expression of EZHIP or H3 K27M in cells promotes similar chromatin profiles: loss of broad H3K27me3 domains but retention at CpG islands. H3K27me3-mediated allosteric activation of PRC2 substantially increases the inhibition potency of EZHIP, providing a mechanism for the observed loss of H3K27me3 spreading. |
In vitro PRC2 catalytic inhibition assay; chromatin profiling (ChIP-seq); cell-based overexpression with H3K27me3 readout; mechanistic dissection of allosteric stimulation |
Nature communications |
High |
31086175
|
| 2019 |
EZHIP (termed CATACOMB) is a subunit of PRC2 whose interaction with the complex decreases PRC2-dependent H3K27me2/3 deposition. The inhibitory function maps to a short conserved region containing a single methionine residue essential for diminishing H3K27me2/3 levels, analogous to H3K27M. EZHIP expression is regulated through DNA methylation/demethylation, suggesting it acts as an inducible endogenous antagonist of PRC2. Additionally, the study identified JAZF1 as a subunit of NuA4 acetyltransferase complex through biochemical characterization of ESS fusion proteins. |
Biochemical characterization; co-purification; methyltransferase assays; site-directed mutagenesis of the critical methionine; epigenetic regulation analysis |
Science advances |
High |
31281901
|
| 2019 |
In mice, EZHIP is predominantly expressed in gonads. Deletion of Ezhip leads to a global increase in H3K27me2/3 during spermatogenesis and at late stages of oocyte maturation. EZHIP limits PRC2 enzymatic activity and lessens the interaction between the core PRC2 complex and its accessory subunits, but does not interfere with PRC2 recruitment to chromatin. Loss of EZHIP is associated with reduced follicles in aging females and strongly impaired fertility in Ezhip−/− females. |
Mouse knockout (Ezhip deletion); ChIP-seq for H3K27me2/3; biochemical analysis of PRC2 complex composition; reproductive phenotyping |
Nature communications |
High |
31451685
|
| 2020 |
EZHIP and H3 K27M preferentially interact with PRC2 that is allosterically activated by H3K27me3 (at CpG islands) and impede its spreading in trans. Expression of human EZHIP in Drosophila melanogaster reduces H3K27me3 through a conserved mechanism, demonstrating mechanistic conservation. H3 K27M oncohistones reduce H3K27me3 in trans, independently of their incorporation into chromatin. |
Biochemical PRC2 binding assays; ChIP-seq in Drosophila expressing human EZHIP; in vivo allosteric activation experiments; domain-specific interaction mapping |
Molecular cell |
High |
33049227
|
| 2020 |
Elevated CXorf67 (EZHIP) expression suppresses homologous recombination (HR)-mediated DNA repair by interacting with PALB2 and inhibiting the PALB2-BRCA2 interaction, thereby blocking HR repair. Tumor cells with high CXorf67 expression show increased sensitivity to PARP inhibitors. |
Co-immunoprecipitation demonstrating CXorf67-PALB2 interaction; HR repair assays; PARP inhibitor sensitivity assays in cells with high vs. low CXorf67 expression |
Cancer cell |
Medium |
33186520
|
| 2013 |
CXorf67 (EZHIP) is identified as the gene on chromosome X involved in a novel reciprocal t(X;17)(p11.2;q21.33) translocation generating the MBTD1-CXorf67 fusion transcript in low-grade endometrial stromal sarcoma, validated by RNA sequencing, RT-PCR, Sanger sequencing, and FISH. |
Whole-transcriptome paired-end RNA sequencing; FISH; banding cytogenetics; RT-PCR and Sanger sequencing |
International journal of cancer |
Medium |
23959973
|
| 2025 |
In growing oocytes, PRC2 binds both classic Polycomb targets and noncanonical H3K27me3 domains; EZHIP co-binds with PRC2 and restricts its activity. Knockout of maternal Ezhip causes hyperactive PRC2 that promiscuously deposits H3K27me3 genome-wide, overwriting H3K27me3 memories at noncanonical imprinted genes, causing paradoxical derepression of H3K27me3 targets, defective X chromosome inactivation, diluted chromatin PRC2, and attenuated H3K27me3 restoration at Polycomb targets after implantation. |
Ezhip knockout in mice; ChIP-seq for H3K27me3 and PRC2 binding; allele-specific analysis of imprinting; X-chromosome inactivation assays |
Cell stem cell |
High |
41118764
|
| 2026 |
Maternal deletion of Ezhip initially increases the asymmetric distribution of H3K27me3 between parental genomes at the zygotic stage, but subsequently impairs H3K27me3-dependent imprinting and mitigates X-chromosome inactivation in pre-implantation embryos. EZHIP protein, translated from the maternal mRNA pool, is present during the first cell divisions post-fertilization and limits PRC2 enzymatic activity; in its absence, the H3K27me3 landscape is both expanded and flattened, abolishing parental genome asymmetry. |
Maternal-specific Ezhip knockout in mice; allele-specific H3K27me3 ChIP-seq; immunofluorescence of early embryos; assessment of X-chromosome inactivation |
Nature communications |
High |
41535302
|
| 2025 |
In a Drosophila model with tissue-specific expression of EZHIP and H3 K27M, a targeted RNAi screen identified genetic modifiers whose knockdown suppressed developmental phenotypes caused by PRC2 inhibition. Strong suppressors included Trithorax-group proteins Ash1 and Trx, the PR-DUB complex member Asx, and the nucleoporin Nup153; suppression correlated with reduced expression of genes aberrantly activated following PRC2 inhibition, placing EZHIP's oncogenic mechanism in the context of conserved chromatin-regulatory pathways. |
Drosophila tissue-specific expression of human EZHIP; targeted RNAi screen; gene expression analysis (RNA-seq); genetic epistasis |
bioRxivpreprint |
Medium |
40502002
|
| 2025 |
EZHIP expression in human-derived neural models potentiates neuronal-like gene programs associated with synaptic function and represses methionine and polyamine metabolism, indicating functions beyond PRC2 inhibition. Additionally, EZHIP mutations occur in H3K27M-positive DMG, showing that mutant EZHIP can be co-expressed with the H3K27M oncohistone contrary to earlier assumptions. |
EZHIP expression in human neural models; transcriptomics (RNA-seq); metabolomics; mutational landscape analysis of pediatric brain tumors |
Acta neuropathologica communications |
Medium |
41204377
|
| 2025 |
Gain- and loss-of-function experiments demonstrate that EZHIP expression drives oncogenic activity in osteosarcoma in vitro and in vivo, reducing H3K27me3 deposition, reactivating developmental pathways, and impeding mesenchymal progenitor differentiation toward smooth muscle lineage at the expense of other fates. |
Gain- and loss-of-function (EZHIP overexpression and knockdown); in vitro and in vivo tumor models; ChIP-seq for H3K27me3; differentiation assays; transcriptomics |
Nature communications |
Medium |
40695784
|
| 2026 |
In silico prediction combined with high-throughput fluorescence-based screening identified that AMPK pathway activation (via biguanides) robustly reduces EZHIP protein abundance and restores H3K27me3 levels independently of cytotoxicity, while PKC activation increases EZHIP protein abundance. This establishes AMPK and PKC as opposing regulators of EZHIP protein stability through post-translational signaling. |
In silico PTM prediction (PhosphoSitePlus, NetPhos 3.1); HEK293T reporter cell line expressing HA-RFP-EZHIP; fluorescence-based high-throughput screening; western blot; LC50 analysis |
Brain tumor research and treatment |
Low |
42120298
|