ORAI3 (ORAI calcium release-activated calcium modulator 3)
2014-06-01 Jessy Hasna , Nazim Benzerdjeb , Malika Faouzi , Anne-Sophie Ay , Philippe Kischel , Frédéric Hague , Henri Sevestre , Ahmed Ahidouch , Halima Ouadid-Ahidouch AffiliationDNA/RNA
Description
ORAI3 is encoded by the gene TMEM142C (HUGO Gene Nomenclature Committee). The ORAI3 gene is located on chromosome 16 in the p11.2.
Transcription
Size of ORAI3 transcript: 2.2 kb; NCBI ORAI3 mRNA model: NM_152288.
All three ORAI isoforms are widely expressed at the mRNA level and can be incorporated into the plasma membrane when ectopically expressed. Broad expression of ORAI3 transcripts has been shown by Northern blot analysis: ORAI3 transcripts are expressed in heart, brain, kidney, thymus, lung, spleen, skeletal muscle, small intestine, as well as in primary aortic endothelial cells and bone marrow derived mast cells (Gwack et al., 2007). ORAI3 appears to be the only family member that is strongly expressed at the RNA level in brain. (ORAI2 transcripts are prominent in kidney, lung, and spleen (Gwack et al., 2007)).
All three ORAI isoforms are widely expressed at the mRNA level and can be incorporated into the plasma membrane when ectopically expressed. Broad expression of ORAI3 transcripts has been shown by Northern blot analysis: ORAI3 transcripts are expressed in heart, brain, kidney, thymus, lung, spleen, skeletal muscle, small intestine, as well as in primary aortic endothelial cells and bone marrow derived mast cells (Gwack et al., 2007). ORAI3 appears to be the only family member that is strongly expressed at the RNA level in brain. (ORAI2 transcripts are prominent in kidney, lung, and spleen (Gwack et al., 2007)).
Transcripts expression
In immune cells, transcripts taken from isolated primary CD3+/CD4+ cells (Th-lymphocytes), CD3+/CD8+ cells (Tc-lymphocytes), CD19+ cells (B-lymphocytes) and BMMC showed that ORAI3 expression is readily detectable in Th-, Tc-, and B- lymphocytes and BMMC (Gross et al., 2007).
mRNA expression in normal tissues has been assessed by different techniques (microarrays, RNAseq, SAGE). Microarrays analyses show that ORAI3 is overexpressed in prostate, lung, monocytes and whole blood (http://biogps.org/#goto=genereport&id=93129, with overexpression defined as 3 times the mean expression observed in the 83 tissues or cells tested in this study). ORAI3 mRNA expression is least important in pancreas, brain (especially the occipital lobe) and T cells (CD4+ as well as CD8+).
Proteins
Schematic representation of ORAI protein structure and organization. Domains of human ORAI1, 2 and 3. P: proline-rich region, R: arginin-rich region, R/K: arginine-lysine-rich region, TM: transmembrane domain, CC: coiled-coil domain (Derler et al., 2012).
Description
Description of the protein sequence.
Molecular weight: 31499 Da.
Sequence length: 295 amino acids.
ORAI3 is a plasma membrane protein containing four transmembrane domains with intracellular N- and C-termini. ORAI3 contains a binding domain for calmodulin in its N-terminus, and a coiled-coil domain for protein interaction in its C-terminus.
Examination of the overall protein sequence of ORAI3 reveals high percentage of homology with the family members: 63.2% with ORAI1 and 66.4% with ORAI2 (60.3% between ORAI1 and ORAI2). These homology percentages increase when the comparison concerns the transmembrane domains: 93.8% with both ORAI1 and ORAI2, (92.5% between ORAI1 and ORAI2) (Feske et al., 2006; Hewavitharana et al., 2007). The pore-forming transmembrane domains of all three ORAI proteins show a high degree (~82%) of conservation.
The amino acid sequence of ORAI3 shows marked differences from its isoforms, particularly in the regions outside of the essential pore-forming domains, which might explain its unique properties and the differences with other isoforms in the modes of regulation and modulation from its isoforms (Shuttleworth, 2012).
The sequence identities between ORAI3 and ORAI1 in the cytosolic N- and C-termini are 34% and 46%, respectively, and is 21% in the extracellular loop between transmembrane domains 3 and 4 (Shuttleworth, 2012).
The N-terminus of ORAI3 comprises ~65 amino acids and has no clusters of prolines and arginines seen in ORAI1 (N-terminus domain containing ~90 amino acids and rich in clusters of prolines and arginines) (Takahashi et al., 2007; Frischauf et al., 2008). ORAI3 has a second extracellular loop linking transmembrane domains 3 and 4 which is longer than that of ORAI1 and ORAI2 (~72 amino acids in ORAI3 compared to only 38 amino acids in ORAI1). ORAI3 has a cluster of 22 positively charged amino acid residues immediately prior to the first transmembrane region which is fully conserved among all three ORAI channels (H44-R66 in ORAI3 and H69-R91 in ORAI1), and has three conserved glutamates located at the C-terminus to which is attributed the fast Ca2+-dependent inactivation of ORAI3 (Lee et al., 2009).
The ORAI3 N-terminus appears critical for switching a store-operated channel to an exclusively arachidonate regulated channel (Thompson et al., 2010).
The residues E81 and E165 in the transmembrane domains 1 and 3, and E85, D87 and E89 in the extracellular 1-2 loop are critical determinants of a high Ca2+ selectivity. Other studies using a cysteine-scanning mutagenesis approach in ORAI3 revealed that Ca2+ selectivity was exclusively determined by the E81 residue alone (McNally et al., 2009).
Replacing the N-terminal cytosolic domain of ORAI3 with the corresponding domain of ORAI1 doubles the magnitude of the measured store-operated Ca2+ currents, whilst the reverse exchange virtually eliminates all currents. N-terminal deletion experiments narrow the critical region essential for the activation of ORAI3 to amino acids 42-62 (Lis et al., 2010). The appearance of significant store-operated currents dependes on a single specific lysine residue K60 in ORAI3, the conservation of this residue in ORAI1 and ORAI3 cannot explain the differences in the magnitude of store-operated Ca2+ currents between these two ORAI family members. N-terminal deletions of residues between W51 and Y55 significantly increase store-operated ORAI3-dependent currents (Bergsmann et al., 2011). The only sequence difference between ORAI1 and ORAI3 in this region is the substitution of a lysine in ORAI1 for an arginine at position 53 in ORAI3.
ORAI3 lacks C195, a reactive cysteine present in ORAI1 that serve as a detection system primarily for changes in the extracellular oxidative environment, and contains two additional cysteines within the extracellular loop between TM3 and TM4. The absence of C195 in ORAI3 makes it resistant to H2O2-inactivation, since pre-incubation with H2O2 of ORAI1/STIM1 expressing cells (HEK; T cells) inhibits activation of ORAI1, but not of ORAI3, and reinsertion of C195 within ORAI3 renders ORAI3 channels redox sensitive (Bogeski et al., 2010).
Molecular weight: 31499 Da.
Sequence length: 295 amino acids.
ORAI3 is a plasma membrane protein containing four transmembrane domains with intracellular N- and C-termini. ORAI3 contains a binding domain for calmodulin in its N-terminus, and a coiled-coil domain for protein interaction in its C-terminus.
Examination of the overall protein sequence of ORAI3 reveals high percentage of homology with the family members: 63.2% with ORAI1 and 66.4% with ORAI2 (60.3% between ORAI1 and ORAI2). These homology percentages increase when the comparison concerns the transmembrane domains: 93.8% with both ORAI1 and ORAI2, (92.5% between ORAI1 and ORAI2) (Feske et al., 2006; Hewavitharana et al., 2007). The pore-forming transmembrane domains of all three ORAI proteins show a high degree (~82%) of conservation.
The amino acid sequence of ORAI3 shows marked differences from its isoforms, particularly in the regions outside of the essential pore-forming domains, which might explain its unique properties and the differences with other isoforms in the modes of regulation and modulation from its isoforms (Shuttleworth, 2012).
The sequence identities between ORAI3 and ORAI1 in the cytosolic N- and C-termini are 34% and 46%, respectively, and is 21% in the extracellular loop between transmembrane domains 3 and 4 (Shuttleworth, 2012).
The N-terminus of ORAI3 comprises ~65 amino acids and has no clusters of prolines and arginines seen in ORAI1 (N-terminus domain containing ~90 amino acids and rich in clusters of prolines and arginines) (Takahashi et al., 2007; Frischauf et al., 2008). ORAI3 has a second extracellular loop linking transmembrane domains 3 and 4 which is longer than that of ORAI1 and ORAI2 (~72 amino acids in ORAI3 compared to only 38 amino acids in ORAI1). ORAI3 has a cluster of 22 positively charged amino acid residues immediately prior to the first transmembrane region which is fully conserved among all three ORAI channels (H44-R66 in ORAI3 and H69-R91 in ORAI1), and has three conserved glutamates located at the C-terminus to which is attributed the fast Ca2+-dependent inactivation of ORAI3 (Lee et al., 2009).
The ORAI3 N-terminus appears critical for switching a store-operated channel to an exclusively arachidonate regulated channel (Thompson et al., 2010).
The residues E81 and E165 in the transmembrane domains 1 and 3, and E85, D87 and E89 in the extracellular 1-2 loop are critical determinants of a high Ca2+ selectivity. Other studies using a cysteine-scanning mutagenesis approach in ORAI3 revealed that Ca2+ selectivity was exclusively determined by the E81 residue alone (McNally et al., 2009).
Replacing the N-terminal cytosolic domain of ORAI3 with the corresponding domain of ORAI1 doubles the magnitude of the measured store-operated Ca2+ currents, whilst the reverse exchange virtually eliminates all currents. N-terminal deletion experiments narrow the critical region essential for the activation of ORAI3 to amino acids 42-62 (Lis et al., 2010). The appearance of significant store-operated currents dependes on a single specific lysine residue K60 in ORAI3, the conservation of this residue in ORAI1 and ORAI3 cannot explain the differences in the magnitude of store-operated Ca2+ currents between these two ORAI family members. N-terminal deletions of residues between W51 and Y55 significantly increase store-operated ORAI3-dependent currents (Bergsmann et al., 2011). The only sequence difference between ORAI1 and ORAI3 in this region is the substitution of a lysine in ORAI1 for an arginine at position 53 in ORAI3.
ORAI3 lacks C195, a reactive cysteine present in ORAI1 that serve as a detection system primarily for changes in the extracellular oxidative environment, and contains two additional cysteines within the extracellular loop between TM3 and TM4. The absence of C195 in ORAI3 makes it resistant to H2O2-inactivation, since pre-incubation with H2O2 of ORAI1/STIM1 expressing cells (HEK; T cells) inhibits activation of ORAI1, but not of ORAI3, and reinsertion of C195 within ORAI3 renders ORAI3 channels redox sensitive (Bogeski et al., 2010).
Post-translational modifications of the protein
Glycosylation:
Unlike ORAI1, ORAI3 does not have a glycosylation site on the asparagine residue (N223) situated between the transmembrane domains TM3 et TM4 (Frischauf et al., 2008; Prakriya et al., 2006).
ORAI1 has a putative N-glycosylation motif (NVS) in its extracellular loop between predicted transmembrane segments 3 and 4. This motif is absent in ORAI2 and 3 (Gwack et al., 2007). ORAI3 migration properties do not change by tunicamycin treatment. Indeed, HEK293 cells stably transfected with FLAG-tagged ORAI and treated with 2μg/ml tunicamycin, showed that ORAI3 migrated at positions close to their predicted molecular masse (32.5 kDa).
Phosphorylation:
Since ORAI3 is a tetraspanning plasma membrane protein, it contains three intracellular regions that can potentially be phosphorylated by intracellular protein kinases: the N-terminus, an intracellular loop between transmembrane domains 2 and 3, and the C-terminus, each intracellular region potentially contains one or more phosphorylation sites. Ser-27 and Ser-30 have been identified as the main phosphorylation sites in ORAI1 within its N-terminus. They are conserved throughout evolution in all mammalian ORAI1 proteins. Mutations at these phosphorylation sites increase store-operated Ca2+ entry (SOCE) and CRAC current suggesting that ORAI1 phosphorylation at these residues by protein kinase C (PKC) suppresses SOCE and CRAC channel activation. However, Ser-27 and Ser-30 are not present in ORAI2 and ORAI3.
A phosphorylation of ORAI3 peptide has been revealed by a phosphoproteome analysis of human liver cells (Sui et al., 2008). This phosphorylation site is located in the C-terminus of ORAI3 on a tyrosine residue (Y278). Experimental ORAI3 phosphorylation has also been demonstrated in HEK293 cells (Kawasaki et al., 2010).
To examine in vivo PKC-mediated phosphorylation, HEK293 cells expressing FLAG-tagged ORAI were incubated with 32P monosodium phosphate, and then stimulated with thapsigargin in the presence of extracellular Ca2+. Thapsigargin mobilizes Ca2+ from the ER and the extracellular space and activates Ca2+/DAG-dependent PKC isoforms. ORAI1 phosphorylation is enhanced in response to thapsigargin. The levels of ORAI3 phosphorylation have been less than half of that observed for ORAI1 (Kawasaki et al., 2010).
Other phosphorylation sites on ORAI3 were predicted by NetPhos2.0: 13 serine sites (S20, S45, S50, S57, S64, S65, S68, S86, S191, S203, S213, S214 and S20), 3 threonine sites (T26, T183 and T190) and 2 tyrosine sites (Y146 and Y278).
ORAI3 protein sequence of amino acids. ORAI3 protein (1 .. 295) has four helical transmembrane domains: T1 (63 .. 82) (20 amino acids), T2 (95 .. 115) (21 amino acids), T3 (157 .. 177) (21 amino acids), T4 (244 .. 264) (21 amino acids).
Expression
ORAI3 is only expressed in mammals (Cai, 2007). ORAI3 seems to be ubiquitously expressed in human (http://www.proteinatlas.org/ENSG00000175938/tissue), and mouse, showing a minor presence in skeletal muscle, spleen and colon (Cordeiro and Strauss, 2011; Gao et al., 2010; Gross et al., 2007). More specifically, ORAI3 expression has been reported in brain, heart, kidney, testis, intestine, placenta, lung (Gwack et al., 2007; Motiani et al., 2013a), vascular smooth muscle cells (Trebak, 2012), airway smooth muscle in human (Peel et al., 2008) and macrophages. ORAI3 mRNA is usually much less expressed compared to ORAI1 in cells of lymphoid origin. ORAI1, ORAI2, and ORAI3 are expressed at similar levels in rat microglia (Hoth and Niemeyer, 2013).
Localisation
ORAI3 localizes to the plasma membrane and functions as a Ca2+-selective ion channel (Feske et al., 2006; Vig et al., 2006; Zhang et al., 2006; Prakriya et al., 2006). This has been confirmed by immunocytochemistry of tagged proteins expressed in Jurkat T cells and in HEK293 cells. All three ORAI isoforms are expressed and localized at or near the plasma membrane, with little or no overlap with the ER marker ERP72. This localization was not grossly altered after store depletion with thapsigargin (Gwack et al., 2007). During meiosis, ORAI proteins get internalized into intracellular vesicles and store-operated currents are suppressed (Yu et al., 2009).
Function
In SOC channels:
ORAI3 presents a single putative channel pore and has a role as a store-operated Ca2+ (SOC) channel. SOC channels are the major route for Ca2+ entry in non-excitable cells, and they include ORAI channels characterized by high selectivity for Ca2+ over monovalent cations, low single-channel conductance (
ORAI3 presents a single putative channel pore and has a role as a store-operated Ca2+ (SOC) channel. SOC channels are the major route for Ca2+ entry in non-excitable cells, and they include ORAI channels characterized by high selectivity for Ca2+ over monovalent cations, low single-channel conductance (
Homology
ORAI3 (encoding gene: MGC13024 located on chromosome 16) has two human homologs: ORAI1 (FLJ14466, chromosome 12) and ORAI2 (C7orf19, chromosome 7) (Feske et al., 2006). ORAI3 made an evolutionary appearance in mammals, evolving from ORAI1 rather than ORAI2 (Cai, 2007) and manifesting conductances that display unique features in their gating, selectivity, regulation and mode of activation (Shuttleworth, 2012).
ORAI3 is the newest ORAI family member in the evolutionary tree (Shuttleworth, 2012). Orthologous ORAI3 genes are found in the following species: chimpanzee (98.98% homology), dog (92.20% homology), cow (90.51% homology), rat (89.83% homology), mice (88.48% homology).
ORAI3 is the newest ORAI family member in the evolutionary tree (Shuttleworth, 2012). Orthologous ORAI3 genes are found in the following species: chimpanzee (98.98% homology), dog (92.20% homology), cow (90.51% homology), rat (89.83% homology), mice (88.48% homology).
Mutations
Note
Understanding of the role of ORAI1, and indeed its initial identification, came from the study of patients carrying functionally critical mutations in this gene. To date, no equivalent identification of patients bearing similar mutations in ORAI3 have been identified. (Diseases associated to absence of ORAI2, ORAI3 or STIM2 function have not been identified in human yet).
Implicated in
Entity name
Breast cancer
Note
ORAI3 channels are reported to be highly expressed in breast cancer (BC) tissues and breast cancer cell lines MCF-7 and T47D compared to adjacent non cancerous tissues and non cancerous cell lines, respectively (Faouzi et al., 2011). They are also shown to be involved in proliferation, cell cycle progression and survival of BC cells by regulating the G1 phase and G1/S transition regulatory proteins. Thus, ORAI3 knockdown by specific siRNA inhibits cell proliferation, arrests cell cycle progression in G1 phase, and increases apoptosis in these cells (Faouzi et al., 2011). This phenotype is associated with a reduction in CDK4 and CDK2 (cyclin-dependent kinases) and cyclin E and cyclin D1 expression, an accumulation of p21Waf1/Cip1 (a cyclin-dependent kinase inhibitor) and p53 (a tumor-suppressor protein) together with an increase of Bax/Bcl-2 ratio. Interestingly, these effects seem to be specific to cancer cells, since down-regulation of ORAI3 channels does not affect either cell proliferation or cell survival of normal breast cells. Annexin V and 7-AAD double staining and analysis of the anti-apoptotic protein Bcl-2 to the pro-apoptotic protein Bax ratio revealed that the induced cell mortality by ORAI3 knockdown was mainly apoptotic as demonstrated by the increased percentage of Annexin V-positive cells and the increased Bax/Bcl-2.
The same study showed that ORAI3 contributes to Ca2+ influx in BC cells where both Store Operated Calcium Entry (SOCE) amplitude and resting [Ca2+]i decreased significantly with ORAI3 knockdown. The authors concluded that the ORAI3 involvement in cell proliferation/survival and cell cycle progression may be at least partially linked to the calcium influx through the channels since the reduction of external calcium concentration [Ca2+]o to 0.2 mM decreases significantly BC cell proliferation (Faouzi et al., 2011).
A subsequent study highlighted a correlation between ORAI3 and the oncogene c-myc expression in tumor tissues and in BC cell lines: ORAI3 and c-myc were over-expressed in 70% and 80% cases respectively. Expression of c-myc, as assessed by RT-qPCR, is higher in the MCF-7 cancer cell line than in the non-cancerous MCF-10A cell line. A similar over-expression pattern was shown for ORAI3 in these cell lines (Faouzi et al., 2013). ORAI3 down-regulation reduces both c-myc expression and activity levels exclusively in BC cells, whereas ORAI1 (one of the two mammalian homologs to ORAI3) induced an upregulation of c-myc mRNA. The involvement of c-myc in the ORAI3 signaling was demonstrated when silencing c-myc resulted in closely-similar and non-additive effects to the ones induced by ORAI3 downregulation: decreased cell proliferation, cell cycle arrest with a significant accumulation of the cells in the G0/G1 phase, increased cell mortality (Faouzi et al., 2013).
Authors showed that ORAI3 channels affect c-myc, most likely via the MAP Kinase pathway, as demonstrated by decreased phosphorylation levels of extracellular signal-regulated kinases 1 and 2 (ERK1/ERK2) after ORAI3 downregulation (Faouzi et al., 2013).
Parallel studies also reported that ORAI3 mediates SOCE in estrogen-receptor-positive (ER+) BC cell lines (Motiani et al., 2010), whereas in estrogen-receptor-negative (ER-) BC cell lines, SOCE is mediated by ORAI1. This study was the first to describe SOCE and endogenous calcium release-activated currents (CRAC) that are mediated by native ORAI3 channels and highlights a potential connection between estrogen receptor alpha (ERα) and ORAI3 (Motiani et al., 2010). Authors then reported that knockdown of ERα decreases ORAI3 expression level leading to a decrease in ORAI3-mediated SOCE and CRAC current, while activation of ERα increased ORAI3 expression and SOCE in MCF7 cells (Motiani et al., 2013b). Consistently with the above cited studies, ORAI3 knockdown inhibits SOCE-dependent phosphorylation of both ERK1/2 and focal adhesion kinase (FAK). It also decreases the transcriptional activity of nuclear factor of activated T-cells (NFAT), which was associated with decreased cell growth and Matrigel invasion of ER+ MCF7 cells in contrast to ER- MDA-MB231 cells where no effects were observed (Motiani et al., 2013b).
The same study showed that ORAI3 contributes to Ca2+ influx in BC cells where both Store Operated Calcium Entry (SOCE) amplitude and resting [Ca2+]i decreased significantly with ORAI3 knockdown. The authors concluded that the ORAI3 involvement in cell proliferation/survival and cell cycle progression may be at least partially linked to the calcium influx through the channels since the reduction of external calcium concentration [Ca2+]o to 0.2 mM decreases significantly BC cell proliferation (Faouzi et al., 2011).
A subsequent study highlighted a correlation between ORAI3 and the oncogene c-myc expression in tumor tissues and in BC cell lines: ORAI3 and c-myc were over-expressed in 70% and 80% cases respectively. Expression of c-myc, as assessed by RT-qPCR, is higher in the MCF-7 cancer cell line than in the non-cancerous MCF-10A cell line. A similar over-expression pattern was shown for ORAI3 in these cell lines (Faouzi et al., 2013). ORAI3 down-regulation reduces both c-myc expression and activity levels exclusively in BC cells, whereas ORAI1 (one of the two mammalian homologs to ORAI3) induced an upregulation of c-myc mRNA. The involvement of c-myc in the ORAI3 signaling was demonstrated when silencing c-myc resulted in closely-similar and non-additive effects to the ones induced by ORAI3 downregulation: decreased cell proliferation, cell cycle arrest with a significant accumulation of the cells in the G0/G1 phase, increased cell mortality (Faouzi et al., 2013).
Authors showed that ORAI3 channels affect c-myc, most likely via the MAP Kinase pathway, as demonstrated by decreased phosphorylation levels of extracellular signal-regulated kinases 1 and 2 (ERK1/ERK2) after ORAI3 downregulation (Faouzi et al., 2013).
Parallel studies also reported that ORAI3 mediates SOCE in estrogen-receptor-positive (ER+) BC cell lines (Motiani et al., 2010), whereas in estrogen-receptor-negative (ER-) BC cell lines, SOCE is mediated by ORAI1. This study was the first to describe SOCE and endogenous calcium release-activated currents (CRAC) that are mediated by native ORAI3 channels and highlights a potential connection between estrogen receptor alpha (ERα) and ORAI3 (Motiani et al., 2010). Authors then reported that knockdown of ERα decreases ORAI3 expression level leading to a decrease in ORAI3-mediated SOCE and CRAC current, while activation of ERα increased ORAI3 expression and SOCE in MCF7 cells (Motiani et al., 2013b). Consistently with the above cited studies, ORAI3 knockdown inhibits SOCE-dependent phosphorylation of both ERK1/2 and focal adhesion kinase (FAK). It also decreases the transcriptional activity of nuclear factor of activated T-cells (NFAT), which was associated with decreased cell growth and Matrigel invasion of ER+ MCF7 cells in contrast to ER- MDA-MB231 cells where no effects were observed (Motiani et al., 2013b).
Entity name
Lung cancer
Note
An overexpression of ORAI3 was observed in 66.7% of human tumor samples as compared to the human non-tumoral samples (40/60) as revealed by immunohistochemistry. The 60 lung adenocarcinomas were classified according to grading system proposed by Yoshizawa et al. 2011 (low, intermediate and high grades). The ORAI3 staining score is reported to be highly expressed in higher tumor grade (high grade; n= 16) as compared to low tumor grades (Ay et al., 2013).
ORAI3 is also expressed in non small cell lung carcinoma cells (NSCLCC) such as NCI-H23, NCI-H460, A549 and Calu-1. In NCI-H23 and NCI-H460 cells, ORAI3 is a major actor of Store Operated Calcium Entry (Ay et al., 2013). Ay et al. (2013) demonstrated that ORAI3 is involved in NSCLCC proliferation. Indeed, ORAI3 inhibition induces a strong decrease in NSCLCC proliferation, accumulating cells in G0/G1 phase of the cell cycle. This accumulation in G0/G1 phase is associated with a decrease in Cyclin D1/cdk4 and Cyclin E/cdk2 proteins level. No effect is observed on apoptosis. The same study demonstrated that SOCE induces Akt phosphorylation in NSCLCC and ORAI3 inhibition decreases this activation demonstrating ORAI3 can promote proliferation through SOCE by activating Akt pathway. They also showed that neither ORAI1 nor ORAI2 are involved in SOCE in NSCLC cell lines, suggesting that ORAI3 is the main component of SOCE in those cells (Ay et al., 2013).
The same type of mechanism is observed with TRPC1 in NSCLCC. Indeed Tajeddine and Gailly (2012) have demonstrated that TRPC1 is involved in G1/S transition in A549 NSCLC cell line through SOCE. They showed that cell cycle arrest after TRPC1 inhibition induces a decrease in EGFR activation and subsequent signaling (PI3K/Akt, MAPK).
Those two studies suggest that SOCE is an important mechanism in proliferation of NSCLCC. Indeed, EGFR signaling is overactivated in NSCLCC either by constitutive activation of EGFR or K-Ras mutation. ORAI3, able to activate this pathway, hence can be a potential target for anti-cancer drug.
ORAI3 is also expressed in non small cell lung carcinoma cells (NSCLCC) such as NCI-H23, NCI-H460, A549 and Calu-1. In NCI-H23 and NCI-H460 cells, ORAI3 is a major actor of Store Operated Calcium Entry (Ay et al., 2013). Ay et al. (2013) demonstrated that ORAI3 is involved in NSCLCC proliferation. Indeed, ORAI3 inhibition induces a strong decrease in NSCLCC proliferation, accumulating cells in G0/G1 phase of the cell cycle. This accumulation in G0/G1 phase is associated with a decrease in Cyclin D1/cdk4 and Cyclin E/cdk2 proteins level. No effect is observed on apoptosis. The same study demonstrated that SOCE induces Akt phosphorylation in NSCLCC and ORAI3 inhibition decreases this activation demonstrating ORAI3 can promote proliferation through SOCE by activating Akt pathway. They also showed that neither ORAI1 nor ORAI2 are involved in SOCE in NSCLC cell lines, suggesting that ORAI3 is the main component of SOCE in those cells (Ay et al., 2013).
The same type of mechanism is observed with TRPC1 in NSCLCC. Indeed Tajeddine and Gailly (2012) have demonstrated that TRPC1 is involved in G1/S transition in A549 NSCLC cell line through SOCE. They showed that cell cycle arrest after TRPC1 inhibition induces a decrease in EGFR activation and subsequent signaling (PI3K/Akt, MAPK).
Those two studies suggest that SOCE is an important mechanism in proliferation of NSCLCC. Indeed, EGFR signaling is overactivated in NSCLCC either by constitutive activation of EGFR or K-Ras mutation. ORAI3, able to activate this pathway, hence can be a potential target for anti-cancer drug.
Entity name
Myeloid leukemia
Note
The mRNA levels of ORAI3 in both human leukemia and human myeloma tipifarnib-sensitive cell lines were significantly higher than in the tipifarnib-insensitive human myeloma cells. Tipifarnib is a new apoptotic agent that inhibits farnesyltransferase responsible for the transfer of a farnesyl group to Ras protein. Tipifarnib activates ORAI3-mediated SOC leading to [Ca2+]i increase. Moreover, ORAI3 functional expression was higher in 2-APB-sensitive leukemia and myeloid cells as compared to 2-APB-insensitive myeloid cells (Yanamadra et al., 2011). These results suggest that Tipifarnib-resistant cells express less ORAI3 ORAI3 conferring protection against apoptotic effect of Tipifarnib (Yanamadra et al., 2011).
Entity name
Prostate cancer
Note
ORAI3 mRNA expression levels are significantly reduced in tumours when compared to non-tumour tissues from 13 prostate cancer patients. mRNA expression levels of ORAI3 are decreased in both androgen-sensitive human prostate adenocarcinoma cell line (LNCaP) and androgen-insensitive prostate cancer cell line (DU145), when compared to human prostate epithelial cells from healthy tissue. The pharmacological effects of 2-APB on CRAC channels in prostate cancer cells differ from those in human prostate epithelial cells, and siRNA based knock-down experiments indicate changed ORAI3 channel levels are underlying the altered pharmacological profile (Holzmann et al., 2013).
Article Bibliography
| Pubmed ID | Last Year | Title | Authors |
|---|---|---|---|
| 24733836 | 2014 | State-dependent block of Orai3 TM1 and TM3 cysteine mutants: insights into 2-APB activation. | Amcheslavsky A et al |
| 24058448 | 2013 | Orai3 constitutes a native store-operated calcium entry that regulates non small cell lung adenocarcinoma cell proliferation. | Ay AS et al |
| 17075073 | 2006 | Coupling of STIM1 to store-operated Ca2+ entry through its constitutive and inducible movement in the endoplasmic reticulum. | Baba Y et al |
| 21724845 | 2011 | Molecular determinants within N terminus of Orai3 protein that control channel activation and gating. | Bergsmann J et al |
| 12220621 | 2002 | Inhibition of the type 1 inositol 1,4,5-trisphosphate receptor by 2-aminoethoxydiphenylborate. | Bilmen JG et al |
| 20107038 | 2010 | Essential role for STIM1/Orai1-mediated calcium influx in PDGF-induced smooth muscle migration. | Bisaillon JM et al |
| 20354224 | 2010 | Differential redox regulation of ORAI ion channels: a mechanism to tune cellular calcium signaling. | Bogeski I et al |
| 17400243 | 2007 | Molecular evolution and structural analysis of the Ca(2+) release-activated Ca(2+) channel subunit, Orai. | Cai X et al |
| 15175387 | 2004 | 2-aminoethoxydiphenyl borate activates and sensitizes the heat-gated ion channel TRPV3. | Chung MK et al |
| 20607548 | 2011 | Expression of Orai genes and I(CRAC) activation in the human retinal pigment epithelium. | Cordeiro S et al |
| 19088073 | 2009 | STIM1- and Orai1-dependent store-operated calcium entry regulates human myoblast differentiation. | Darbellay B et al |
| 18487204 | 2008 | Complex actions of 2-aminoethyldiphenyl borate on store-operated calcium entry. | DeHaven WI et al |
| 19622747 | 2009 | A Ca2(+ )release-activated Ca2(+) (CRAC) modulatory domain (CMD) within STIM1 mediates fast Ca2(+)-dependent inactivation of ORAI1 channels. | Derler I et al |
| 22453951 | 2012 | Structure, regulation and biophysics of I(CRAC), STIM/Orai1. | Derler I et al |
| 19754892 | 2009 | Mechanistic view on domains mediating STIM1-Orai coupling. | Fahrner M et al |
| 20683915 | 2011 | Down-regulation of Orai3 arrests cell-cycle progression and induces apoptosis in breast cancer cells but not in normal breast epithelial cells. | Faouzi M et al |
| 23266555 | 2013 | ORAI3 silencing alters cell proliferation and cell cycle progression via c-myc pathway in breast cancer cells. | Faouzi M et al |
| 20887894 | 2010 | Store-independent activation of Orai1 by SPCA2 in mammary tumors. | Feng M et al |
| 16582901 | 2006 | A mutation in Orai1 causes immune deficiency by abrogating CRAC channel function. | Feske S et al |
| 21220423 | 2011 | Cooperativeness of Orai cytosolic domains tunes subtype-specific gating. | Frischauf I et al |
| 20630587 | 2010 | Calcium-activated K(+) channel (K(Ca)3.1) activity during Ca(2+) store depletion and store-operated Ca(2+) entry in human macrophages. | Gao YD et al |
| 11171105 | 2001 | Evidence that 2-aminoethyl diphenylborate is a novel inhibitor of store-operated Ca2+ channels in liver cells, and acts through a mechanism which does not involve inositol trisphosphate receptors. | Gregory RB et al |
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Other Information
Locus ID:
NCBI: 93129
MIM: 610930
HGNC: 28185
Ensembl: ENSG00000175938
Variants:
dbSNP: 93129
ClinVar: 93129
TCGA: ENSG00000175938
COSMIC: ORAI3
RNA/Proteins
Expression (GTEx)
Pathways
| Pathway | Source | External ID |
|---|---|---|
| Calcium signaling pathway | KEGG | ko04020 |
| Calcium signaling pathway | KEGG | hsa04020 |
Protein levels (Protein atlas)
References
| Pubmed ID | Year | Title | Citations |
|---|---|---|---|
| 37759448 | 2023 | Orai3 Calcium Channel Contributes to Oral/Oropharyngeal Cancer Stemness through the Elevation of ID1 Expression. | 0 |
| 37759448 | 2023 | Orai3 Calcium Channel Contributes to Oral/Oropharyngeal Cancer Stemness through the Elevation of ID1 Expression. | 0 |
| 34626722 | 2022 | High ORAI3 expression correlates with good prognosis in human muscle-invasive bladder cancer. | 2 |
| 34626722 | 2022 | High ORAI3 expression correlates with good prognosis in human muscle-invasive bladder cancer. | 2 |
| 33798603 | 2021 | ORAI3 silencing alters cell proliferation and promotes mitotic catastrophe and apoptosis in pancreatic adenocarcinoma. | 8 |
| 34768857 | 2021 | Role of Orai3 in the Pathophysiology of Cancer. | 8 |
| 34943998 | 2021 | Orai3 Calcium Channel Regulates Breast Cancer Cell Migration through Calcium-Dependent and -Independent Mechanisms. | 11 |
| 33798603 | 2021 | ORAI3 silencing alters cell proliferation and promotes mitotic catastrophe and apoptosis in pancreatic adenocarcinoma. | 8 |
| 34768857 | 2021 | Role of Orai3 in the Pathophysiology of Cancer. | 8 |
| 34943998 | 2021 | Orai3 Calcium Channel Regulates Breast Cancer Cell Migration through Calcium-Dependent and -Independent Mechanisms. | 11 |
| 32896813 | 2020 | Distinct pharmacological profiles of ORAI1, ORAI2, and ORAI3 channels. | 53 |
| 32896813 | 2020 | Distinct pharmacological profiles of ORAI1, ORAI2, and ORAI3 channels. | 53 |
| 30943377 | 2019 | Bronchial Epithelial Calcium Metabolism Impairment in Smokers and Chronic Obstructive Pulmonary Disease. Decreased ORAI3 Signaling. | 16 |
| 30943377 | 2019 | Bronchial Epithelial Calcium Metabolism Impairment in Smokers and Chronic Obstructive Pulmonary Disease. Decreased ORAI3 Signaling. | 16 |
| 29237733 | 2018 | Communication between N terminus and loop2 tunes Orai activation. | 30 |
Citation
Jessy Hasna ; Nazim Benzerdjeb ; Malika Faouzi ; Anne-Sophie Ay ; Philippe Kischel ; Frédéric Hague ; Henri Sevestre ; Ahmed Ahidouch ; Halima Ouadid-Ahidouch
ORAI3 (ORAI calcium release-activated calcium modulator 3)
Atlas Genet Cytogenet Oncol Haematol. 2014-06-01
Online version: http://atlasgeneticsoncology.org/gene/51589/orai3-(orai-calcium-release-activated-calcium-modulator-3)
