NOL3 (nucleolar protein 3 (apoptosis repressor with CARD domain))

2009-05-01 Gloria Kung , Wendy McKimpson , Richard N Kitsis Affiliation

Department of Medicine, Department of Cell Biology, Montefiore-Einstein Center for Cardiovascular Research, Albert Einstein Cancer Center, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY 10461 USA

Identity

HGNC

NOL3

LOCATION

16q22.1

LOCUSID

8996

ALIAS

ARC,FCM,MYOCL1,MYP,NOP,NOP30

FUSION GENES

Show Gene Fusions

DNA/RNA

The NOL3 gene is located on the long arm of human chromosome 16. The gene consists of 4 small exons (exons denoted above as thick boxes) and 3 small introns. The translational start site is in exon 2. Alternative splicing occurs between exons 2 and 3. This involves two splice donors separated by 10 nucleotides in exon 2 connecting to a single splice acceptor in exon 3 (Stoss et al., 1999). Because the separation between the splice donors, 10 nucleotides, is not an exact multiple of 3, alternative splicing results in two open reading frames distal to the splice acceptor. Because of this frame shift, the C-terminus of the two encoded proteins differ as do their stop codons, each of which is in exon 4. One transcript is translated into ARC (Apoptosis Repressor with CARD (Caspase Recruitment Domain)) (Koseki et al., 1998). MYP is an earlier name for ARC that is no longer in use (Geertman et al., 1996). The other transcript encodes a putative protein called NOP30 (Nucleolar Protein of 30 kD). ARC and putative NOP30 proteins share a common N-terminus containing the CARD. Their C-termini differ, however, with ARC containing multiple P/E repeats (acidic) and putative NOP30 containing R/S repeats (basic). While ARC transcripts are present in a variety of human and mouse cell types, NOP30 transcripts are present in human, but not mouse (L. Wu and R. Kitsis, unpublished). Endogenous ARC protein resides in the cytoplasm and nucleoplasm of certain human and mouse cell types (discussed below). In contrast, the existence of endogenous NOP30 protein has not been demonstrated in any cell type of any species. When the cDNA encoding NOP30 is exogenously expressed, the encoded protein is in the nucleolus and nucleoplasm (Stoss et al., 1999).

Description

The NOL3 gene is located on human chromosome 16q21-23. The gene contains 4 exons and 3 introns spanning 1757 bp.

Transcription

The coordinate of the first nucleotide of exon 1 is 65,765,371 bp from pter, and that of the last nucleotide of exon 4 is 65,767,127 bp. Alternative splicing takes place between exons 2 and 3. In exon 2, the splice donor of the NOP30 transcript is 10 bp upstream of the splice donor of the ARC transcript. Both transcripts use a common splice acceptor in exon 3.

Proteins

Note

The start of translation is in exon 2 (prior to the alternative splice donors). Alternative splicing causes a frame shift resulting in transcripts encoding proteins with different C-termini and separate stop codons in exon 4. The stop codon for ARC is 43 bp upstream of that of NOP30.

Alternatively spliced transcripts of NOL3 lead to two different proteins, ARC (blue) and NOP30 (red). These proteins each contain an N-terminal CARD (first 95 amino acids identical), but have different C-termini. The C-terminus of ARC is rich in prolines and glutamic acids, whereas the C-terminus of NOP30 is rich in serines and arginines.

Description

Human ARC protein contains 208 amino acids with Mr 22,629 Da. The protein usually runs at a slower mobility on SDS-PAGE most likely due to the enrichment of proline residues in the C-terminal domain. NOP30 contains 219 amino acids with Mr 24,327 Da.

Expression

Under normal conditions, ARC mRNA and protein is present predominantly in cardiac myocytes, skeletal myocytes, and neurons (Koseki et al., 1998; Abmayr et al., 2004; Geertman et al., 1996; Engidawork et al., 2001). ARC protein is also markedly increased in primary human epithelial cancers of the breast, colon, ovary, and cervix (Mercier et al., 2005; Mercier et al., 2008). NOP30 transcripts are present in some human cell types but have not been detected in mouse cells. Endogenous NOP30 protein has not been demonstrated in cells of any species.

Localisation

Endogenous ARC protein is present in the cytoplasm and nucleoplasm (Mercier et al., 2005). As above, the localization of endogenous NOP30 protein has not been investigated. Exogenously expressed NOP30 protein localizes in the nucleolus and nucleoplasm.

Function

The function of endogenous NOP30 is not known. Exogenous NOP30 interacts with SFRS9/SRp30C and NPM1 and may influence splicing (Stoss et al., 1999).
ARC is an endogenous inhibitor of apoptosis that is unique in its ability to antagonize both the extrinsic (death receptor) and the intrinsic (mitochondria/ER) death pathways (Nam et al., 2004; Gustafsson et al., 2004; Koseki et al., 1998). ARC inhibits the extrinsic pathway by interfering with DISC (Death Inducing Signaling Complex) formation. This is accomplished by the direct interaction of the ARC CARD with the death domains (DD) of Fas and FADD, and with the death effector domain (DED) of procaspase-8. These death-fold interactions are novel in that they are heterotypic in contrast to the usual homotypic death-fold interactions. ARC inhibits the intrinsic pathway through at least two mechanisms. First, the direct interaction between the ARC CARD and the C-terminus of Bax inhibits death stimulus-induced Bax conformational activation and translocation to the mitochondria. Second, direct interaction between the ARC C-terminal domain with the p53 tetramerization domain inhibits p53 tetramerization (Foo et al., PNAS, 2007). This, in turn, disables p53 transcriptional function and exposes a p53 nuclear export signal that relocates p53 to the cytoplasm.

Nothing is known about the regulation of NOP30.
The regulation of ARC is complex. ARC protein abundance decreases rapidly and dramatically in response to hypoxia and oxidative stress (e.g. ischemia-reperfusion) (Ekhterae et al., 1999; Neuss et al., 2001; Nam et al., 2007). These decreases result from increased degradation of ARC protein via the ubiquitin-proteasomal pathway (Nam et al., 2007). The E3 ligase MDM2 may play a role in ARC degradation in this scenario (Foo et al., JBC, 2007), but this role is probably indirect (L. Wu and R. Kitsis, unpublished data). Decreases in ARC protein abundance in response to hypoxia appear to be regulated by p53 repression of nol3 transcription (Li et al., 2008). Apart from ARC protein abundance, the activity of ARC is also regulated post-translationally: dephosphorylation of threonine 149 decreases the anti-apoptotic activity of ARC (Tan et al., 2008).

Regulation of the extrinsic (death receptor) and intrinsic (mitochondria/ER) apoptosis pathways by ARC. Not shown are ARC interactions with and regulation of p53.

Homology

ARC is highly conserved among mammals. There is approximately 85% identity both at the amino acid and the nucleotide level among human, rat, mouse, dog, and bovine ARC. Interestingly, an ARC homolog has yet to be identified in Danio rerio, Drosophila melanogaster, or Caenorhabditis elegans.

Implicated in

Entity name

Epithelial cancers

Disease

Increased levels of ARC protein have been observed in the epithelium of primary human breast, colon, ovarian, and cervical cancers (Mercier et al., 2005; Mercier et al., 2008). Increased levels of both ARC RNA and protein have been observed in renal cell carcinoma (Heikaus et al., 2008).

Prognosis

ARC overexpression in a breast cancer cell line increases resistance to chemotherapy and radiation (Mercier et al., 2005; Wang et al., 2009). In a melanoma cell line, ARC overexpression causes increased resistance to endoplasmic reticulum stress-induced caspase-8 activation (Chen et al., 2008).

Entity name

Myocardial infarction, myocardial ischemia-reperfusion

Prognosis

ARC plays an important role in regulating heart muscle damage during myocardial infarction. Endogenous ARC protein undergoes rapid proteasomal degradation during myocardial ischemia-reperfusion (Nam et al., 2007). This decrease in ARC abundance is causally linked with the subsequent cell death (Nam et al., 2004). Accordingly, transgenic overexpression of ARC in vivo decreases the size of myocardial infarctions (Gustafsson et al., 2002; Pyo et al., 2008; S. Jha and R. Kitsis, unpublished data). As would be predicted, knockout of ARC has been reported to result in larger infarcts (Donath et al., 2006). However, the aforementioned knockout studies were performed on only small numbers of mice on a mixed genetic background. Subsequent knockout studies involving large numbers of mice on several pure genetic backgrounds have not demonstrated larger infarcts in ARC-/- mice subjected to ischemia-reperfusion (J. Saurabh, S. Y. Ji, and R. Kitsis, unpublished data). This is probably due to the dramatic rapid degradation of ARC protein during reperfusion even in wild type mice (see above).

Entity name

Heart failure

Prognosis

ARC protein levels decrease during heart failure. Moreover, knockout of ARC exacerbates pathological cardiac remodeling in response to hemodynamic overload, a model of heart failure (Donath et al., 2006).

Entity name

Neuropathology (several individual entities)

Prognosis

The protein level of ARC is increased in the frontal cortex of patients with Alzheimers disease (Engidawork et al., 2001). During ischemic injury of the brain, there is a decrease in ARC protein in hippocampal neurons (Hong et al., 2003). Other studies have also shown that caloric restriction increases expression of ARC in the brain (Shelke et al., 2003).

Breakpoints

Note

Not known.

Bibliography

Pubmed ID	Last Year	Title	Authors
14645204	2004	Characterization of ARC, apoptosis repressor interacting with CARD, in normal and dystrophin-deficient skeletal muscle.	Abmayr S et al
18245485	2008	Inhibition of endoplasmic reticulum stress-induced apoptosis of melanoma cells by the ARC protein.	Chen LH et al
16505176	2006	Apoptosis repressor with caspase recruitment domain is required for cardioprotection in response to biomechanical and ischemic stress.	Donath S et al
10590251	1999	ARC inhibits cytochrome c release from mitochondria and protects against hypoxia-induced apoptosis in heart-derived H9c2 cells.	Ekhterae D et al
11178964	2001	Alteration of caspases and apoptosis-related proteins in brains of patients with Alzheimer's disease.	Engidawork E et al
17142834	2007	Ubiquitination and degradation of the anti-apoptotic protein ARC by MDM2.	Foo RS et al
18087040	2007	Regulation of p53 tetramerization and nuclear export by ARC.	Foo RS et al
8634331	1996	Cloning and characterization of cDNAs for novel proteins with glutamic acid-proline dipeptide tandem repeats.	Geertman R et al
15004034	2004	Apoptosis repressor with caspase recruitment domain protects against cell death by interfering with Bax activation.	Gustafsson AB et al
18516683	2008	Caspase-8 and its inhibitors in RCCs in vivo: the prominent role of ARC.	Heikaus S et al
12753927	2003	Down-regulation of ARC contributes to vulnerability of hippocampal neurons to ischemia/hypoxia.	Hong YM et al
9560245	1998	ARC, an inhibitor of apoptosis expressed in skeletal muscle and heart that interacts selectively with caspases.	Koseki T et al
17998337	2008	p53 initiates apoptosis by transcriptionally targeting the antiapoptotic protein ARC.	Li YZ et al
18469522	2008	ARC (apoptosis repressor with caspase recruitment domain) is a novel marker of human colon cancer.	Mercier I et al
17142452	2007	The apoptosis inhibitor ARC undergoes ubiquitin-proteasomal-mediated degradation in response to death stimuli: identification of a degradation-resistant mutant.	Nam YJ et al
11438535	2001	The apoptotic regulatory protein ARC (apoptosis repressor with caspase recruitment domain) prevents oxidant stress-mediated cell death by preserving mitochondrial function.	Neuss M et al
18782777	2008	Protection of cardiomyocytes from ischemic/hypoxic cell death via Drbp1 and pMe2GlyDH in cardio-specific ARC transgenic mice.	Pyo JO et al
12514107	2003	Lifelong caloric restriction increases expression of apoptosis repressor with a caspase recruitment domain (ARC) in the brain.	Shelke RR et al
10196175	1999	Alternative splicing determines the intracellular localization of the novel nuclear protein Nop30 and its interaction with the splicing factor SRp30c.	Stoss O et al
19001025	2008	Novel cardiac apoptotic pathway: the dephosphorylation of apoptosis repressor with caspase recruitment domain by calcineurin.	Tan WQ et al
19147562	2009	Apoptosis repressor with caspase recruitment domain contributes to chemotherapy resistance by abolishing mitochondrial fission mediated by dynamin-related protein-1.	Wang JX et al

Other Information

Locus ID:

NCBI: 8996
MIM: 605235
HGNC: 7869
Ensembl: ENSG00000140939

Variants:

dbSNP: 8996
ClinVar: 8996
TCGA: ENSG00000140939
COSMIC: NOL3

RNA/Proteins

Gene ID	Transcript ID	Uniprot
ENSG00000140939	ENST00000268605	O60936
ENSG00000140939	ENST00000268605	Q5TZN6
ENSG00000140939	ENST00000563258	J3QLT7
ENSG00000140939	ENST00000563439	H3BM67
ENSG00000140939	ENST00000564860	J3QLS5
ENSG00000140939	ENST00000564992	H3BUN4
ENSG00000140939	ENST00000565560	H3BUP2
ENSG00000140939	ENST00000566871	H3BQJ5

Expression (GTEx)

Adipose - Subcutaneous

Adipose - Visceral

Adrenal Gland

Artery - Aorta

Artery - Tibial

Bladder

Brain - Amygdala

Brain - Anterior cingulate cortex

Brain - Caudate

Brain - Cerebellar Hemisphere

Brain - Cerebellum

Brain - Cortex

Brain - Frontal Cortex

Brain - Hippocampus

Brain - Hypothalamus

Brain - Nucleus accumbens

Brain - Putamen

Brain - Spinal cord

Brain - Substantia nigra

Breast - Mammary Tissue

Cells - Cultured fibroblasts

Cells - EBV - transformed lymphocytes

Cervix - Ectocervix

Cervix - Endocervix

Colon - Sigmoid

Colon - Transverse

Esophagus - Gastroesophageal Junction

Esophagus - Mucosa

Esophagus - Muscularis

Fallopian Tube

Heart - Atrial Appendage

Heart - Left Ventricle

Kidney - Cortex

Kidney - Medulla

Liver

Lung

Minor Salivary Gland

Muscle - Skeletal

Nerve - Tibial

Ovary

Pancreas

Pituitary

Prostate

Skin - Not Sun Exposed

Skin - Sun Exposed

Small Intestine - Terminal Ileum

Spleen

Stomach

Testis

Thyroid

Uterus

Vagina

Whole Blood

Protein levels (Protein atlas)

Not detected

Low

Medium

High

References

Pubmed ID	Year	Title	Citations
19913121	2009	Gene-centric association signals for lipids and apolipoproteins identified via the HumanCVD BeadChip.	85
15383280	2004	Inhibition of both the extrinsic and intrinsic death pathways through nonhomotypic death-fold interactions.	53
15383280	2004	Inhibition of both the extrinsic and intrinsic death pathways through nonhomotypic death-fold interactions.	53
15383280	2004	Inhibition of both the extrinsic and intrinsic death pathways through nonhomotypic death-fold interactions.	53
18087040	2007	Regulation of p53 tetramerization and nuclear export by ARC.	37
15861191	2005	ARC, an apoptosis suppressor limited to terminally differentiated cells, is induced in human breast cancer and confers chemo- and radiation-resistance.	32
18469522	2008	ARC (apoptosis repressor with caspase recruitment domain) is a novel marker of human colon cancer.	30
24763054	2014	MicroRNA-185 regulates chemotherapeutic sensitivity in gastric cancer by targeting apoptosis repressor with caspase recruitment domain.	30
16505176	2006	Apoptosis repressor with caspase recruitment domain is required for cardioprotection in response to biomechanical and ischemic stress.	29
22037876	2011	Apoptosis inhibitor ARC promotes breast tumorigenesis, metastasis, and chemoresistance.	26

Citation

Gloria Kung ; Wendy McKimpson ; Richard N Kitsis

NOL3 (nucleolar protein 3 (apoptosis repressor with CARD domain))

Atlas Genet Cytogenet Oncol Haematol. 2009-05-01

Online version: http://atlasgeneticsoncology.org/gene/41552/nol3