| 2006 |
The PHD domain of ING2 binds specifically and with high affinity to trimethylated and dimethylated histone H3 lysine 4 (H3K4me3/me2), functioning as an effector module for this histone mark. In response to DNA damage, this interaction stabilizes the mSin3a-HDAC1 repressive complex at promoters of proliferation genes, constituting a mechanism by which H3K4me3 drives active gene repression. Mutations disrupting H3K4me3 binding abolish ING2-mediated cellular responses to genotoxic insults. |
Biochemical binding assays, ChIP, loss-of-function mutagenesis, co-immunoprecipitation, cellular DNA damage response assays |
Nature |
High |
16728974
|
| 2006 |
Crystal structure (2.0 Å) of the mouse ING2 PHD finger in complex with H3K4me3 peptide revealed that the trimethylammonium group of Lys4 is recognized by aromatic residues Y215 and W238 in a deep binding groove, with additional hydrogen-bonding contacts to Ala1, Arg2, Thr3, and Thr6 of the histone tail. Substitution of binding-site residues disrupts H3K4me3 interaction in vitro and impairs ING2-induced apoptosis in vivo. |
X-ray crystallography (2.0 Å resolution), in vitro binding assays, site-directed mutagenesis, apoptosis assays |
Nature |
High |
16728977
|
| 2003 |
The PHD finger of ING2 binds phosphoinositides in vitro, including phosphatidylinositol 5-phosphate (PtdIns(5)P). The ING2 PHD finger interacts with PtdIns(5)P in vivo in the nucleus, and this interaction regulates the ability of ING2 to activate p53 and p53-dependent apoptotic pathways in response to DNA damage. |
In vitro lipid-binding assays, in vivo interaction studies, p53 transcriptional assays, apoptosis assays |
Cell |
High |
12859901
|
| 2001 |
ING2 (p33ING2) negatively regulates cell growth in a p53-dependent manner and enhances p53 transcriptional activity. ING2 expression increases acetylation of p53 at Lys-382, indicating ING2 promotes p53 acetylation as part of its tumor suppressive function. ING2 is induced by DNA-damaging agents etoposide and neocarzinostatin. |
Cell proliferation assays, p53 transactivation reporter assays, Western blot for p53 acetylation, siRNA knockdown, overexpression |
Proceedings of the National Academy of Sciences of the United States of America |
Medium |
11481424
|
| 2005 |
ING2 forms a complex with p53 and the histone acetyltransferase p300, enhances the interaction between p53 and p300, and acts as a cofactor for p300-mediated p53 acetylation. ING2-mediated p300-dependent p53 acetylation triggers replicative senescence; overexpression induces senescence in young fibroblasts in a p53-dependent manner, and siRNA knockdown of ING2 delays senescence onset. |
Co-immunoprecipitation, siRNA knockdown, overexpression, senescence assays, colocalization studies |
Molecular and cellular biology |
Medium |
16024799
|
| 2010 |
Treatment with the HDAC inhibitor SAHA causes dissociation of the ING2 subunit (via its PHD finger) from the Sin3 deacetylase complex. Loss of ING2 from the complex disrupts in vivo binding of the Sin3 complex to the p21 promoter, revealing a molecular mechanism by which HDAC inhibitors disrupt deacetylase function at target gene promoters. |
Mass spectrometry-based proteomics, Co-IP, ChIP at p21 promoter, pharmacological treatment with SAHA |
Chemistry & biology |
Medium |
20142042
|
| 2006 |
ING2 enhances nucleotide excision repair of UV-induced DNA damage in a p53-dependent manner. ING2 is required for UV-induced histone H4 acetylation, chromatin relaxation, and recruitment of the damage-recognition protein XPA to photolesions. Knockdown of ING2 completely abolishes NER, demonstrating that physiological ING2 levels are required for this process. |
Host-cell reactivation assay, siRNA knockdown, histone acetylation assays, chromatin accessibility assays, XPA recruitment by immunofluorescence |
Cancer research |
Medium |
16488987
|
| 2006 |
The leucine zipper-like (LZL) motif in the N-terminus of ING2 is critical for DNA repair, apoptosis, and chromatin remodeling after UV irradiation. Deletion of the LZL domain abrogates the association between ING2 and p53, but not between ING2 and p300, indicating ING2 functions as a scaffold mediating p53-p300 interaction. |
Domain deletion mutagenesis, Co-immunoprecipitation, apoptosis assays, NER assays |
FEBS letters |
Medium |
16782091
|
| 2010 |
The HECT-type ubiquitin ligase Smurf1 interacts with ING2 and targets it for polyubiquitination and proteasomal degradation. The catalytic HECT domain of Smurf1 mediates ING2 binding, and the C-terminal PHD domain of ING2 is required for Smurf1-mediated degradation. |
Co-immunoprecipitation, ubiquitination assays, proteasome inhibitor experiments, domain mapping |
FEBS letters |
Medium |
20621832
|
| 2013 |
Nuclear phosphatidylinositol-5-phosphate (PtdIns(5)P) directly associates with ING2 and is required for ING2 occupancy at a subset of genomic target promoters in response to DNA damage. PtdIns(5)P acts as a sub-nuclear trafficking factor stabilizing ING2 at discrete chromatin sites; depletion of PtdIns(5)P attenuates ING2-mediated gene repression at these targets. |
ChIP, PtdIns(5)P depletion, gene expression analysis, lipid-binding assays |
Scientific reports |
Medium |
23823870
|
| 2008 |
ING2 promotes TGF-β-induced transcription and cell cycle arrest. ING2 interacts with the transcriptional modulator SnoN, and together they form a complex with Smad2. Knockdown of SnoN blocks ING2-dependent TGF-β transcription, while SnoN expression augments it, placing ING2 in the TGF-β-Smad signaling pathway as a co-activator. |
Co-immunoprecipitation, RNA interference, overexpression, transcriptional reporter assays, cell cycle analysis |
The Journal of biological chemistry |
Medium |
18334480
|
| 2009 |
ING2 interacts with PCNA (proliferating cell nuclear antigen) and regulates the amount of PCNA associated with chromatin, thereby controlling DNA replication fork progression. siRNA-mediated knockdown of ING2 markedly reduces global replication rate (DNA fiber spreading), causes endoreduplication, and increases sister chromatid exchange frequency, demonstrating a role for ING2 in maintaining genome stability. |
Co-immunoprecipitation, DNA fiber spreading assay, siRNA knockdown, chromatin fractionation, sister chromatid exchange assay |
EMBO reports |
Medium |
19730436
|
| 2010 |
ING2 controls the G1/S transition by regulating p21 expression independently of p53. Knockdown of ING2 accelerates G1-to-S phase progression and decreases p21 levels. This function is specific to ING2 and not shared by its closest homolog ING1. |
siRNA knockdown, cell cycle analysis (FACS), RT-PCR/Western blot for p21, comparative ING1 vs ING2 experiments |
Cell cycle (Georgetown, Tex.) |
Medium |
20890119
|
| 2010 |
ING2 is sumoylated by SUMO1 on lysine 195 both in vitro and in vivo. Sumoylation of ING2 enhances its association with Sin3A and is required for ING2 binding to the promoters of specific target genes (e.g., TMEM71) and for recruitment of the Sin3A/HDAC complex to those promoters to regulate transcription. |
In vitro sumoylation assay, in vivo SUMO modification assay, Co-IP, ChIP, site-directed mutagenesis (K195R) |
Oncogene |
Medium |
20676127
|
| 2008 |
ING2 associates with histone methyltransferase (HMT) activity in vitro and in vivo, methylating histone H3 with a specificity distinct from the MeCP2-recruited HMT. The ING2-associated HMT shows increased activity when H3K9 is already methylated, but reduced activity when H3K4 is mutated or methylated. The C-terminus of ING2 recruits this HMT activity and correlates with its gene silencing function, which is HDAC-independent (resistant to trichostatin A). |
In vitro HMT assay, co-immunoprecipitation, reporter-based silencing assays, domain deletion/mutation analysis, TSA resistance assay |
Biochimica et biophysica acta |
Medium |
18513492
|
| 2010 |
Targeted germline disruption of Ing2 in mice causes male infertility due to defective spermatogenesis with meiotic arrest before pachytene stage, incomplete meiotic recombination, and enhanced apoptosis. Arrested spermatocytes lacked specific HDAC1 accumulation and showed deregulated chromatin acetylation, implicating an ING2/HDAC1/H3K4me3-regulated chromatin modification pathway in spermatogenesis. Ing2-null mice also develop soft-tissue sarcomas, confirming tumor suppressor function in vivo. |
Targeted gene knockout in mice, histology, sperm count/motility assays, immunostaining for HDAC1 and histone acetylation marks, tumor incidence analysis |
PloS one |
High |
21124965
|
| 2009 |
ING2 protein levels increase upon MNNG treatment in an MMR (MLH1)- and c-Abl-dependent manner. MNNG-induced ING2 localizes to the nucleus and associates with p73α. Suppression of ING2 by shRNA decreases MNNG sensitivity and abrogates MNNG-induced stabilization and acetylation of p73α, placing ING2 downstream of MMR/c-Abl and upstream of p73α in the alkylation-induced cell death pathway. |
shRNA knockdown, immunoprecipitation, immunofluorescence, Western blot, kinase inhibitor (STI571) treatment, cell viability assays |
Experimental cell research |
Medium |
19766113
|
| 2007 |
ING2 interacts in vivo with the corepressor Alien (also known as TRIP15/CSN2). The interaction was confirmed in vitro by GST pulldown, indicating direct binding. The binding domain was mapped to a central region of Alien. Co-expression of ING2 enhances Alien-mediated transcriptional silencing. |
SELDI-MS proteomics, co-immunoprecipitation, GST pulldown, transcriptional silencing assays |
Journal of proteome research |
Medium |
17929852
|
| 2016 |
ING2 acts as a corepressor for the androgen receptor (AR): ING2 interacts with AR, hampers AR transcriptional activation, causes growth arrest, and induces cellular senescence in prostate cancer cells. ING2 protein levels are upregulated as a compensatory mechanism when ING1b is knocked down, suggesting a crosstalk between ING1 and ING2 to co-regulate AR signaling. |
Co-immunoprecipitation, siRNA/shRNA knockdown, luciferase reporter assays, cell growth assays, senescence assays, Ing1 knockout mouse tissues |
Journal of molecular medicine (Berlin, Germany) |
Medium |
27305909
|
| 2021 |
ING2 is imported into the inner mitochondrial compartment in a redox-sensitive manner, and this translocation is modulated by 14-3-3η protein expression. Mitochondrial ING2 interacts with mtDNA, and this interaction is mediated by TFAM. Loss of mitochondrial ING2 decreases mitochondrial ROS production and impairs OXPHOS activity, establishing ING2 as a regulator of mitochondrial respiration and metabolic homeostasis. |
Subcellular fractionation, immunofluorescence/confocal microscopy, Co-immunoprecipitation (ING2-TFAM), oxygen consumption rate (OCR) assay, ROS measurement, siRNA knockdown |
Oncogene |
Medium |
34017078
|
| 2021 |
ING2 positively regulates mitochondrial respiration in tubular epithelial cells by controlling the ubiquitination and stability of MRPL12, a mitochondrial transcription factor, thereby modulating mtDNA transcription and expression of mtDNA-encoded respiratory chain components. ING2 overexpression in vivo ameliorates acute ischemic kidney injury. |
Co-immunoprecipitation, ubiquitination assay, oxygen consumption rate assay, PCR/Western blot, immunofluorescence, in vivo kidney-specific overexpression mouse model |
Frontiers in cell and developmental biology |
Medium |
34434929
|
| 2021 |
ING1 and ING2 are recruited to the negative androgen response element (nARE) in the hTERT core promoter in an androgen-dependent manner, and knockdown of ING1 and ING2 blocks AR-mediated repression of hTERT, establishing both as AR co-repressors required for supraphysiological androgen-induced hTERT repression. |
ChIP, siRNA knockdown, luciferase reporter assays, RT-PCR, cancer spheroid models |
Cancers |
Medium |
34439179
|
| 2012 |
ING2 is required for myogenic differentiation of C2C12 cells; RNAi knockdown blocks differentiation into myotubes. Structure-function analysis shows the leucine zipper motif is required for ING2-driven differentiation, while the PHD domain inhibits this function. The Sin3A-HDAC1 complex, which interacts with ING2, is also required for ING2-dependent muscle differentiation. |
RNAi knockdown, overexpression, domain deletion/mutation analysis, myogenic differentiation assay, Co-immunoprecipitation |
PloS one |
Medium |
22808232
|
| 2009 |
ING2 upregulates matrix metalloproteinase 13 (MMP13) expression, and this regulation requires the ING2-HDAC1-mSin3A complex. Co-expression of ING2 with HDAC1 or mSin3A further induces MMP13. ING2 overexpression increases invasive potential of colon cancer cells, establishing a functional link between ING2 chromatin complex activity and MMP13-dependent invasion. |
Microarray, overexpression, siRNA knockdown, EMSA, luciferase assays (NF-κB on ING2 promoter), in vitro invasion assay, RT-PCR/Western blot |
International journal of cancer |
Medium |
19437536
|