ING4 (inhibitor of growth family, member 4)

2010-12-01   Angela Greco , Claudia Miranda 

Dept Experimental Oncology, Molecular Mechanisms Unit, Istituto Nazionale Tumori IRCCS Foundation - via Venezian 1 - 20133 Milan Italy

Identity

HGNC
LOCATION
12p13.31
LOCUSID
ALIAS
my036,p29ING4
FUSION GENES

DNA/RNA

Atlas Image
Figure adapted from Atlas of Genetics and Cytogenetics in Oncology and Haematology.

Description

ING4 belongs to family of highly homologous five members containing PHD domain and has been identified through a computational sequence homology search for expressed tag clones with a PHD finger motif (Shiseki et al., 2003). ING4 gene is located on chromosome 12p13.31 and consists of eight exons encoding a 29-kDa protein expressed in multiple human tissues.

Transcription

Multiple alternatively spliced transcript variants have been observed using different splice sites in the coding region; transcript variants span from 1461 bp to 1313 bp.
Wobble splicing events have been described at exon 4 and 5 boundary. Different splicing variants have been identified (among them -v1, -v2, -v3 and -v4/Δ4AA) involving 12 bp (379-390) and resulting in in frame deletions of one to four aminoacids in NLS (Tsai and Lin, 2006).
Several splicing variants have been described lacking exon 2, 3 and 6 (entirely or in part), and named ING4-ΔEx2, -ΔEx3, -ΔEx6A and -ΔEx6B, respectively (Raho et al., 2007).
Splicing variants have been detected in all tissues analysed, indicating that are not tissue specific (Tsai and Lin, 2006; Raho et al., 2007).
More recently five novel spliced variants of ING4-v1 and -v2 were identified, causing codon frame shift and eventually deletion of NLS or PHD domains. Increased expression of these variants was found in gastric adenocarcinomas compared to normal tissue (Li M et al., 2009).

Proteins

Note

249 aminoacids, 29 kDa protein.
Atlas Image
LZL: leucine zipper-like; NLS1: nuclear localization signal 1; PHD: plant homology domain; NLS2: nuclear localization signal 2.

Description

ING4 protein contains several conserved regions: i) a leucine zipper-like (LZL) domain, probably involved in protein interactions, located at the N-terminus; ii) a functional bipartite nuclear localization signal (NLS1); iii) a C-terminal plant homeo-domain (PHD), a Cys4-His-Cys3 zinc finger motif spanning 50-80 residues, found in many nuclear proteins, such as transcription factors and proteins regulating chromatin structure; iv) a non functional NLS located at the C-terminal end.

Expression

Ubiquitous.

Localisation

p29ING4 is a nuclear protein. It possess a bipartite nuclear localization signal. ING4 splicing variants have been described involving the NLS1 domain; most/all of them retain nuclear localization. Furthermore ING4-v1 is translocated to the nucleolus and such subcellular localization is modulated by two wobble-splicing events at the exon 4-5 boundary, causing displacement from the nucleolus to the nucleus.

Function

ING4 was also isolated through a screening for genes able to suppress loss of contact inhibition, thus suggesting its tumor suppressor role.
ING4 is a nuclear protein participating to a variety of cellular functions, such as apoptosis, cell-cycle regulation, chromatin remodeling, and regulation of gene expression. Several ING4 partners have been described. Similarly to the other ING members, ING4 was described to interact with p53 and to modulate p53 transcriptional activity (Shiseki et al., 2003). The interaction of ING4 with p53 is mediated by the bipartite ING4 nuclear localization signal (NLS) (Zhang et al., 2005) and drives an increase of p53 acetylation at lysine 382 (Shiseki et al., 2003). ING4 is a critical regulator of chromatin acetylation required for gene expression. In particular, ING4 associates with the HAT complex HBO1 and it is required for the majority of histone 4 acetylation and for normal progression through S phase (Doyon et al., 2006; Shi et al., 2006). Recently a critical role for specific recognition of histone H3 trimethylated at lysine 4 (H3K4me3) by the ING4 PHD finger in mediating ING4 gene expression and tumor suppressor functions has been shown (Hung et al., 2009).
ING4 can also function as repressor of factors mediating angiogenesis. It was demonstrated that ING4 plays an inhibitory role on NF-kappaB activity by interaction with p65NF-kappaB and that the lack of inhibition of the NF-kappaB pathway by ING4 results in increased angiogenesis in glioblastomas (Garkavtsev et al., 2004). More recently, it has been described that physiologic levels of ING4 govern innate immunity in mice by regulating the levels of IkappaB and NF-kappaB proteins and the activation of select cytokine promoters (Coles et al., 2010).
ING4 was also described to repress the ability of hypoxia inducible factor (HIF)-1 to activate transcription of its downstream target genes by interacting with the HPH-2 prolyl hydroxylase. Under hypoxic conditions, ING4 may act as an adapter protein recruiting transcriptional repressors to mediate HIF activity (Ozer et al., 2005).
Involvement of ING4 in regulation of apoptosis has been demonstrated in several cellular systems. Its overexpression can induce apoptosis through the downregulation of Bcl-2 and the upregulation of p21 and Bax expression (Shiseki et al., 2003; Yu et al., 2007; Li X et al., 2009b; Cai et al., 2009).

Homology

ING4 protein shares homology with other ING family members with respect to the following regions: i) a leucine zipper-like (LZL) domain, probably involved in interaction with proteins, located at the N-terminus of all the ING proteins except for ING1; ii) a nuclear localization signal (NLS); iii) a C-terminal plant homeo-domain (PHD) involved in chromatin.

Mutations

Note

The following ING4 point mutations have been found in lung adenocarcinoma and small cell lung carcinoma (H23 and H28, respectively) human cancer cell lines.
N214D: it alters ING4 capability of inhibition of proliferation, anchorage independent cell migration reducing protein stability by proteasome mediated degradation.
Y121N: it does not alter ING4 functions (Moreno et al., 2010).

Implicated in

Entity name
Breast cancer
Cytogenetics
Analysis of CGH data revealed that 10-20% of primary breast tumors present deletions in 12p13. The deletions appear to affect only one copy of the gene; no genomic mutations were found in the remaining allele of ING4 (Kim et al., 2004).
Entity name
Head and neck squamous cell carcinoma (HNSCC)
Cytogenetics
LOH of 12p13.
Oncogenesis
Loss of heterozygosity at 12p12-13 region was found in 66% (33/50) of head and neck squamous cell carcinomas by using six highly polymorphic microsatellite markers. No mutations of the ING4 gene were found.
Quantitative real-time RT-PCR analysis demonstrated decreased expression of ING4 mRNA in 76% of primary tumors compared to matched normal samples (Gunduz et al., 2005).
Entity name
Glioma
Oncogenesis
Expression of ING4 is significantly reduced in gliomas as compared with normal human brain tissue, and the extent of reduction correlates with the progression from lower to higher grades of tumours. ING4 regulates brain tumour angiogenesis through transcriptional repression of NF-kB-responsive genes (Garkavtsev et al., 2004).
Entity name
Astrocytoma
Prognosis
A potential of role of ING4 as a biomarker for the prediction of the grade of astrocytic neoplasms has been suggested (Klironomos et al., 2010).
Oncogenesis
Significantly reduced levels of ING4 were observed in human astrocytomas compared to normal brain tissue, suggesting that down-regulation of this protein might be involved in the pathogenesis of human astrocytic tumors. Decreased ING4 expression correlated significantly with tumor progression, with lower expression levels of ING4 observed in cases of high-grade neoplasms. A statistical significant negative correlation between expression of ING4 and expression of nuclear p65 was noticed (Klironomos et al., 2010).
Entity name
Hepatocellular carcinoma (HCC)
Prognosis
Survival and metastasis analysis indicated that HCC patients with lower ING4 expression had poorer overall and disease-free survival than those with high expression. Multivariable Cox regression analysis revealed that the ING4 expression level was an independent factor for prognosis (Fang et al., 2009).
Oncogenesis
The ING4 mRNA and protein levels were significantly lower in HCC than paracarcinomatous liver tissue. ING4 expression level correlates with prognosis and metastatic potential, suggesting that ING4 as a candidate prognostic marker of HCC (Fang et al., 2009).
Entity name
Prognosis
MM patients with high IL-8 production and microvascular density (MVD) have significantly lower ING4 levels compared with those with low IL-8 and MVD.
Oncogenesis
ING4 suppression in MM cells up-regulated IL-8 and OPN under hypoxic conditions, increasing the hypoxia inducible factor-1alpha (HIF-1alpha) activity and its target gene NIP-3 expression. ING4 suppression in MM cells significantly increased vessel formation in vitro, blunted by blocking IL-8 or OPN (Colla et al., 2007).
Entity name
Lung cancer
Oncogenesis
Reduced ING4 nuclear and cytoplasmic expression were both revealed in lung cancer and associated with tumour grade. ING4 expression in the cytoplasm was found higher than in the nucleus in a high percentage of tumors. Nuclear ING4 inhibition correlated with the tumour stage and lymph node metastasis, thus suggesting that ING4 is involved in the initiation and progression of lung cancers (Wang et al., 2010).
Entity name
Gastric cancer
Oncogenesis
ING4 RNA and protein were drastically reduced in stomach adenocarcinoma cell lines and tissues, significantly less in female than male patients. Novel spliced forms of ING4-v1 and -v2 were identified in both normal and tumor tissue; increased expression of the novel spliced variants was observed in tumors; however no correlation with clinical parameters was observed (Li M et al., 2009).

Bibliography

Pubmed IDLast YearTitleAuthors
194304012009Inhibitor of growth 4 is involved in melanomagenesis and induces growth suppression and apoptosis in melanoma cell line M14.Cai L et al
205345382010Inhibitor of growth-4 promotes IkappaB promoter activation to suppress NF-kappaB signaling and innate immunity.Coles AH et al
178486182007The new tumor-suppressor gene inhibitor of growth family member 4 (ING4) regulates the production of proangiogenic molecules by myeloma cells and suppresses hypoxia-inducible factor-1 alpha (HIF-1alpha) activity: involvement in myeloma-induced angiogenesis.Colla S et al
163876532006ING tumor suppressor proteins are critical regulators of chromatin acetylation required for genome expression and perpetuation.Doyon Y et al
192086632009Decreased expression of inhibitor of growth 4 correlated with poor prognosis of hepatocellular carcinoma.Fang F et al
150291972004The candidate tumour suppressor protein ING4 regulates brain tumour growth and angiogenesis.Garkavtsev I et al
159355702005Frequent deletion and down-regulation of ING4, a candidate tumor suppressor gene at 12p13, in head and neck squamous cell carcinomas.Gunduz M et al
191877652009ING4 mediates crosstalk between histone H3 K4 trimethylation and H3 acetylation to attenuate cellular transformation.Hung T et al
155282762004A screen for genes that suppress loss of contact inhibition: identification of ING4 as a candidate tumor suppressor gene in human cancer.Kim S et al
205018482010A dominant mutant allele of the ING4 tumor suppressor found in human cancer cells exacerbates MYC-initiated mouse mammary tumorigenesis.Kim S et al
197752942010Loss of inhibitor of growth (ING-4) is implicated in the pathogenesis and progression of human astrocytomas.Klironomos G et al
194798222009Reduced expression and novel splice variants of ING4 in human gastric adenocarcinoma.Li M et al
195716072009Inhibitor of growth 4 induces growth suppression and apoptosis in glioma U87MG.Li X et al
190345112009Inhibitor of growth 4 induces apoptosis in human lung adenocarcinoma cell line A549 via Bcl-2 family proteins and mitochondria apoptosis pathway.Li X et al
207059532010Functional impact of cancer-associated mutations in the tumor suppressor protein ING4.Moreno A et al
187793152008The ING4 tumor suppressor attenuates NF-kappaB activity at the promoters of target genes.Nozell S et al
160963742005Regulation of HIF by prolyl hydroxylases: recruitment of the candidate tumor suppressor protein ING4.Ozer A et al
158974522005The candidate tumor suppressor ING4 represses activation of the hypoxia inducible factor (HIF).Ozer A et al
200533572010The dimeric structure and the bivalent recognition of H3K4me3 by the tumor suppressor ING4 suggests a mechanism for enhanced targeting of the HBO1 complex to chromatin.Palacios A et al
173256602007Detection of novel mRNA splice variants of human ING4 tumor suppressor gene.Raho G et al
167289742006ING2 PHD domain links histone H3 lysine 4 methylation to active gene repression.Shi X et al
127502542003p29ING4 and p28ING5 bind to p53 and p300, and enhance p53 activity.Shiseki M et al
169203302006Quantitative analysis of wobble splicing indicates that it is not tissue specific.Tsai KW et al
187756962008Two wobble-splicing events affect ING4 protein subnuclear localization and degradation.Tsai KW et al
192505432009Down-regulation of the inhibitor of growth family member 4 (ING4) in different forms of pulmonary fibrosis.Tzouvelekis A et al
169736152006Novel splice variants of ING4 and their possible roles in the regulation of cell growth and motility.Unoki M et al
207161692010Down-regulation of ING4 is associated with initiation and progression of lung cancer.Wang QS et al
179394062007[Ad-ING4 inhibits K562 cell growth].Yu X et al
158829812005Nuclear localization signal of ING4 plays a key role in its binding to p53.Zhang X et al

Other Information

Locus ID:

NCBI: 51147
MIM: 608524
HGNC: 19423
Ensembl: ENSG00000111653

Variants:

dbSNP: 51147
ClinVar: 51147
TCGA: ENSG00000111653
COSMIC: ING4

RNA/Proteins

Gene IDTranscript IDUniprot
ENSG00000111653ENST00000341550Q9UNL4
ENSG00000111653ENST00000396807Q9UNL4
ENSG00000111653ENST00000412586Q9UNL4
ENSG00000111653ENST00000423703Q9UNL4
ENSG00000111653ENST00000444704Q9UNL4
ENSG00000111653ENST00000446105Q9UNL4
ENSG00000111653ENST00000467678R4GNG1
ENSG00000111653ENST00000469749A4KYM5
ENSG00000111653ENST00000479301E9PJ14
ENSG00000111653ENST00000482489H0YEH7
ENSG00000111653ENST00000488381E9PJ14
ENSG00000111653ENST00000493873E9PNE3
ENSG00000111653ENST00000619641A4KYM5

Expression (GTEx)

0
10
20
30
40
50
60
70
80

Pathways

PathwaySourceExternal ID
Chromatin organizationREACTOMER-HSA-4839726
Chromatin modifying enzymesREACTOMER-HSA-3247509
HATs acetylate histonesREACTOMER-HSA-3214847

Protein levels (Protein atlas)

Not detected
Low
Medium
High

References

Pubmed IDYearTitleCitations
150291972004The candidate tumour suppressor protein ING4 regulates brain tumour growth and angiogenesis.101
127502542003p29ING4 and p28ING5 bind to p53 and p300, and enhance p53 activity.94
158974522005The candidate tumor suppressor ING4 represses activation of the hypoxia inducible factor (HIF).51
187793152008The ING4 tumor suppressor attenuates NF-kappaB activity at the promoters of target genes.49
274711082017miR-423-5p contributes to a malignant phenotype and temozolomide chemoresistance in glioblastomas.38
211778152011EBNA3C attenuates the function of p53 through interaction with inhibitor of growth family proteins 4 and 5.36
155282762004A screen for genes that suppress loss of contact inhibition: identification of ING4 as a candidate tumor suppressor gene in human cancer.34
152514302004ING4 induces G2/M cell cycle arrest and enhances the chemosensitivity to DNA-damage agents in HepG2 cells.32
178486182007The new tumor-suppressor gene inhibitor of growth family member 4 (ING4) regulates the production of proangiogenic molecules by myeloma cells and suppresses hypoxia-inducible factor-1 alpha (HIF-1alpha) activity: involvement in myeloma-induced angiogenesis.32
203814592010MicroRNA-650 targets ING4 to promote gastric cancer tumorigenicity.32

Citation

Angela Greco ; Claudia Miranda

ING4 (inhibitor of growth family, member 4)

Atlas Genet Cytogenet Oncol Haematol. 2010-12-01

Online version: http://atlasgeneticsoncology.org/gene/40978/ing4