|Coordinate||3,436,475 bp (GRCm38)|
|Base Change||A ⇒ G (forward strand)|
|Gene Name||DNA cross-link repair 1C|
|Synonym(s)||Artemis, Art, 9930121L06Rik|
|Chromosomal Location||3,424,131-3,464,130 bp (+)|
FUNCTION: This gene encodes a member of the SNM1 family of nucleases and is involved in V(D)J recombination and DNA repair. This protein has single-strand-specific 5'-3' exonuclease activity; it also exhibits endonuclease activity on 5' and 3' overhangs and hairpins. The protein also functions in the regulation of the cell cycle in response to DNA damage. Homozygous knockout mice for this gene exhibit severe combined immunodeficiency with sensitivity to ionizing radiation. Mutations in this gene in humans can cause Athabascan-type severe combined immunodeficiency (SCIDA) and Omenn syndrome. Alternative splicing results in multiple transcript variants encoding different isoforms. [provided by RefSeq, Nov 2014]
PHENOTYPE: Homozygous mutant mice exhibit a combined immunodeficiency phenotype. While immunoglobulin rearrangement is completely blocked in B cells, the block of V(D)J rearrangement in T cells is partial. [provided by MGI curators]
|Amino Acid Change||Histidine changed to Arginine|
|Institutional Source||Beutler Lab|
|Gene Model||predicted gene model for protein(s): [ENSMUSP00000054300] [ENSMUSP00000098031] [ENSMUSP00000100053] [ENSMUSP00000110718]|
AA Change: H115R
|Predicted Effect||probably damaging
PolyPhen 2 Score 1.000 (Sensitivity: 0.00; Specificity: 1.00)
AA Change: H115R
|Predicted Effect||probably damaging
PolyPhen 2 Score 1.000 (Sensitivity: 0.00; Specificity: 1.00)
AA Change: H115R
|Predicted Effect||probably damaging
PolyPhen 2 Score 1.000 (Sensitivity: 0.00; Specificity: 1.00)
|Predicted Effect||probably benign|
|Predicted Effect||probably benign|
|Meta Mutation Damage Score||0.9678|
|Is this an essential gene?||Possibly nonessential (E-score: 0.329)|
|Candidate Explorer Status||CE: excellent candidate; Verification probability: 0.968; ML prob: 0.954; human score: 3.5|
Linkage Analysis Data
|Alleles Listed at MGI|
|Mode of Inheritance||Autosomal Recessive|
|Local Stock||Live Mice|
|Last Updated||2018-06-15 11:54 AM by Diantha La Vine|
|Record Created||2013-07-20 1:15 PM by Kuan-Wen Wang|
The kiwis phenotype was identified among N-ethyl-N-nitrosourea (ENU)-mutagenized G3 mice of the pedigree R0520, some of which showed a decreased frequency of B cells (Figure 1) including B1a (Figure 2) and B2 (Figure 3) cells in the peripheral blood; the frequency of B1b cells was increased (Figure 4). Some mice exhibited a reduced percentage of IgD+ B cells (Figure 5) and a reduced frequency of IgM+ B cells (Figure 6). The mean fluorescence intensity (MFI) of IgD (Figure 7) and B220 (Figure 8) on B cells were diminished.
Some mice exhibited a reduced frequency of T cells (Figure 9) including both CD4+ (Figure 10) and CD8+ T cells (Figure 11) in the peripheral blood. The CD44 MFI was increased on T cells (Figure 12) including both CD4+ (Figure 13) and CD8+ T cells (Figure 14). The frequency of natural killer T (NKT) cells was reduced in the peripheral blood (Figure 15).
Some mice showed an increased frequency of macrophages (Figure 16), neutrophils (Figure 17), natural killer (NK) cells (Figure 18), and CD11c+ dendritic cells (Figure 19), all in the peripheral blood.
The T-dependent antibody responses to ovalbumin administered with aluminum hydroxide (Figure 20) as well as to the T-dependent antigen recombinant Semliki Forest virus (rSFV)-encoded β-galactosidase (rSFV-β-gal) (Figure 21) were diminished. Also, the T-independent antibody response to 4-hydroxy-3-nitrophenylacetyl-Ficoll (NP-Ficoll) was diminished (Figure 22).
|Nature of Mutation|
Whole exome HiSeq sequencing of the G1 grandsire identified 73 mutations. All of the above anomalies were linked by continuous variable mapping to a mutation in Dclre1c: an A to G transition at base pair 3,436,475 (v38) on chromosome 2, or base pair 12,684 in the GenBank genomic region NC_000068. The strongest association was found with a recessive model of linkage to the normalized peripheral blood B2 cell frequency, wherein 5 variant homozygotes departed phenotypically from 10 homozygous reference mice and 16 heterozygous mice with a P value of 1.099 x 10-19 (Figure 23). A substantial semidominant effect was observed in most of the assays but the mutation is preponderantly recessive, and in no assay was a purely dominant effect observed.
The mutation corresponds to residue 431 in the mRNA sequences NM_146114, NM_175683, NM_001110214 within exon 5 of 14, 16, and 15 total exons, respectively.
Genomic numbering corresponds to NC_000068. The mutated nucleotide is indicated in red. The mutation results in a histidine (H) to arginine (R) substitution at position 115 (H115R) in the Artemis protein, and is strongly predicted by Polyphen-2 to cause loss of function (score = 1.00).
|Illustration of Mutations in
Gene & Protein
Dclre1c encodes Artemis, a member of the metallo-β-lactamase protein superfamily (1). The metallo-β-lactamase proteins have two conserved domains: a metallo-β-lactamase domain (amino acids 10-193 in Artemis, SMART) and a β-CASP (metallo-β-lactamase-associated CPSF ARTEMIS SNM1 PSO2) domain (amino acids 239-345 in Artemis, SMART) [Figure 24; (2); reviewed in (3)]. Together, the metallo-β-lactamase and β-CASP domains are designated as the SNM1 domain [reviewed in (4)]. The SNM1 domain comprises the “catalytic core” of Artemis and contains nuclease activity as well as regulates protein-protein interactions including that of Artemis with the Cul4a-DDB1 ubiquitin complex [(5-7); reviewed in (4)].
The metallo-β-lactamase domain is a four-layered β-sandwich with two mixed β-sheets flanked by α-helices (1;8;9). The metallo-β-lactamase domain has five highly conserved sequence motifs that function in metal coordination, substrate binding, and enzymatic activities (8;9). Motif 1 contains an aspartate at the end of two β-strands of the first β-sheet (8;10). Motif 2 contains a HxHxDH sequence (amino acids 33-38 in Artemis); the first two His residues in the motif (His33 and His35) are proposed to participate in metal ion coordination (10). The Asp amino acid in the motif (Asp37) is proposed to function in the hydrolysis reactions; the role of the third His (His38) in the motif is unknown (10). Asp37, His33, His35, His38, His115, and His319 coordinate the two active site metals, while Asp17, Asp136 and/or Asp165 are proposed to form salt bridges to the HxHxDH motif as well as H38 and H33 to stabilize the HxHxDH motif for optimum metal ion interaction (10). Motifs 3 and 5 are comprised of single histidines that are proposed to coordinate metal ions and the binding of negatively charged substrates (10). Motif 4 is a single aspartate that is proposed to participate in hydrolysis reactions (8;9). Asp165 or His319 are proposed to represent motif 5 (2;5). The β-CASP domain is between motif 4 and motif 5 and has an α/β fold with a five-stranded β-sheet surrounded on both sides with α-helices (11). Amino acid 341 determines nucleic acid specificity (i.e., a His at 341 is found in RNA-specific enzymes, while a Val at 341 is found in DNA-specific enzymes (2); Val341 is essential for the interaction of Artemis with DNA (5).
The C-terminal region (CTR; amino acids 346-705) is dispensable for Artemis-mediated hairpin opening during V(D)J recombination (7) and is also required for DNA repair (12;13). Trp489, Phe492, and Ph493 within the CTR are essential for the formation of the Artemis-Ligase IV/XRCC4 complex (14). Ma et al. proposed an inhibitory role for the CTR, which would regulate Artemis function in the absence of DNA double-strand breaks (DSBs) (7). Several sites within the CTR of Artemis are phosphorylated by the phosphatidylinositol-3-OH kinase-like (PIK) kinases DNA-dependent protein kinase catalytic subunit (DNA-PKCS; see the record for clover), ataxia telangiectasia mutated (ATM), and ATM- and Rad3-related (ATR) in response to DNA damage or cellular stress (7;15-18). Three basal (Ser503, Ser516, and Ser645) and 11 DNA-PKcs–mediated phosphorylation sites have been located in the CTR (6;7;19). Ser645 is also phosphorylated by ATM in response to ionizing radiation (IR) (15;17) (15). Mutation of Ser645 to alanine (Ser645Ala) did not affect survival of the mutant cells after exposure to IR, indicating that phosphorylation at Ser645 is not functionally critical (7). Artemis phosphorylation is not required for Artemis-dependent DSB repair and V(D)J recombination (see “Background” section for more information about V(D)J recombination) (6).
DNA DSBs can occur as a result of exposure to external factors such as IR (21) and toxins (e.g., asbestos, silica, and titanium dioxide) (22). Cellular processes such as the generation of reactive oxygen species, the collapse of DNA replication forks (upon recognition of single-stranded breaks by the replication machinery) (23), and, in the case of B and T lymphocytes, immune receptor gene arrangement, also cause DSBs (24-27). Repair of DSBs is required to prevent chromosomal abnormalities and chromosome loss, and thereby maintain genomic stability. If left unrepaired, cell cycle arrest typically occurs, leading to cell death (26;28;29). In addition, instances of cancer can occur after a tumor suppressor gene is inactivated or deleted by a DSB, or when an oncogene is activated or translocated (30;31).
There are three DNA DSB repair pathways: single-strand annealing (SSA), homologous recombination (HR), and nonhomologous end-joining (NHEJ) (32). In SSA, which requires homologous sequences to flank the break site, the nonhomologous DNA ends are removed, leaving overhangs that are aligned and annealed (26). After annealing, DNA synthesis and ligation occur. SSA results in deletion of the region between homologous sequences. Homologous recombination (HR) and nonhomologous end-joining (NHEJ) occur upon recognition of a DNA DSB (Figure 25).
HR is a relatively error-free mechanism for DSB repair that relies on the homologous region on the sister chromatid as a template for DNA synthesis [Figure 25A; (33)]. HR is limited to replicating cells (including early embryonic developmental stages) and can be observed only after DNA replication in the S and G2 phases (34). The endonuclease function of Artemis is essential for HR at heterochromatin during the G2 phase of the cell cycle (35). The function of Artemis in HR is independent of DNA-PKcs (35).
Nonhomologous end-joining (NHEJ)
NHEJ is a rapid repair mechanism that simply ligates broken DNA ends after minimal or no processing (Figure 25B). NHEJ occurs in all phases of the cell cycle and is thought to be the primary DNA repair pathway in mammalian cells (36). NHEJ is prone to generating mutations at the point of ligation, and may result in inversions or translocations if the joined strands were not originally contiguous. For a detailed description of NHEJ, please the record for clover. In NHEJ, autophosphorylation of DNA-PKCS results in release of DNA-PKCS from the DNA ends and accessibility of the termini to enzymes and ligases needed to complete the DSB repair (e.g. Artemis, DNA polymerase X family members and the DNA ligase IV-XRCC4 dimer) (24;33;37). Artemis and DNA-PKCS form a complex with endonuclease activity that cleaves 5’ and 3’ overhangs during NHEJ, removes 3′ phosphoglycolate termini, and opens hairpins generated by the RAG complex (see the record for maladaptive for information about Rag1 and the record for snowcock for information about Rag2) during V(D)J recombination (6;38-40). In order to activate its nuclease activity, Artemis is phosphorylated by DNA-PKCS (39). Autophosphorylation of DNA-PKcs, and subsequent phosphorylation of Artemis, as well as the presence of Ku70/80 facilitate the hairpin and 5’ and 3’ overhang endonuclease activity of Artemis at sites of single-stranded to double-stranded DNA junctions (6;7;39-41). At a 5’ overhang, the Artemis/DNA-PKcs complex preferentially cuts at the double-stranded/single-stranded junction (39;42). At a 3’ overhang, Artemis preferentially cuts 4 nucleotides into the single-stranded overhang from the double/single-stranded DNA junction (43). Artemis opens hairpins typically two nucleotides past the tip of a perfect hairpin (39). In the absence of DNA-PKcs, Artemis displays 5’-3’ exonucleolease activity (7;10;17).
Immunoglobulin and T cell receptor loci consist of linear arrays of gene segments that require combinatorial assembly to form functional coding sequences (Figure 26). During lymphoid cell development, V-J or V-D-J segments of Ig or TCR loci are joined by the process of V(D)J recombination to generate a variable region exon, which is subsequently linked to the C region gene by RNA splicing. Ultimately, pre-B cells and thymocytes can survive to maturity only if they successfully carry out V(D)J recombination that will give them in-frame Ig and TCR chains, to be assembled into the final B cell receptor (BCR) and TCR complexes. For more information on V(D)J recombination, please see the record for maladaptive. RAG1/2 cleave the DNA within the synaptic complex, yielding the cleaved signal complex. Hairpin structures on the coding ends are nicked by DNA-PKcs and Artemis and the DNA DSBs induced by the RAG proteins is repaired using the NHEJ pathway (39;44-46).
Class Switch Recombination (CSR)
Class switch recombination (CSR) facilities the production of antibodies of different isotypes in mature B cells during a humoral immune response (47;48). CSR is a recombination reaction that occurs between paired DSBs in immunoglobulin heavy chain (Igh) switch regions (S-regions) that flank Igh constant regions (49). The S-regions contain a repetitive sequence that can serve as a substrate for proximal microhomology-mediated intra-switch repair by C-NHEJ (50). During CSR, activation-induced cytidine deaminase (AID) converts cytosines into uracils at the S-region (51). The excision of uracils from both DNA strands results in staggered DNA breaks at donor and acceptor switch regions (51). The Igh locus lesions are detected as DSBs by the MRN [MRE11 (meiotic recombination 11)–Rad50–NBS1 (Nijmegen breakage syndrome 1)] complex, which leads to phosphorylation of H2AX, the recruitment of 53BP1(see the record for lentil) to the Igh locus, and eventual end joining by C- or A-NHEJ (49). For more information on CSR, please see the lentil page. During CSR, DNA-PKcs and Artemis prevent chromosomal translocations by promoting end-joining of IgH locus DSBs to other IgH locus DSBs on the same chromosome (52). The endonuclease function of Artemis is required for the C-NHEJ pathway in CSR at 3’ or 5’ overhangs (53). Examination of S recombinational junctions from DCLRE1C-deficient B cells determined that Sμ–Sα junctions lacked direct end-joining and shifted towards the use of a microhomology-based end-joining pathway (53). At the Sμ–Sγ junctions, there was an increase of sequential switching from IgM, through one IgG subclass, to a different IgG subclass (53). The Sγ–Sγ junctions showed long microhomologies (53).
Cell cycle regulation
Artemis regulates recovery from the G2 checkpoint in response to IR through regulation of cyclin B/Cdk1 activation by retaining Cdk1/cyclin B at the centrosome and inhibiting its nuclear import during prophase (15-18). Mutation of Ser516 and Ser645 in Artemis to alanines (Ser516Ala and Ser645Ala) prevented phosphorylation of Artemis by ATM and resulted in a slower recovery from the G2/M checkpoint (18;54). Artemis is also involved in S phase checkpoint recovery in response to replication fork blocking lesions in an phosphorylation-dependent manner (54). At the S phase checkpoint recovery, Artemis interacts with SCFFbw7 to mediate the degradation of cyclin E via the SCFFbw7 E3 ligase complex (54). Phosphorylation of Artemis at Ser516 and Ser645 by ATR in response to replication stress (e.g., UVC, aphidicolin, and hydroxyurea) promotes the interaction with Fbw7 subsequently facilitating cyclin E turnover and S phase recovery (54). Artemis interacts with the tumor suppressor p27 during the G1 phase of the cell cycle and is required for the ubiquitination and degradation of p27 by the Cul4A-DDB1 complex, which is required cell cycle progression at the G1-S and G0 to S transitions (55).
Artemis is required for normal proliferative control of multipotent mesenchymal stem/progenitor cells (MSCs), especially after exposure to cytostress stimuli (56). Dclre1c deficiency resulted in chromosomal damage as well as enhanced resistance and proliferative potential in primary MSCs after stress (56).
Artemis is a negative regulator of p53 in response to oxidative stress in primary cells and cancer cell lines (57). Loss of Artemis expression results in phosphorylation and stabilization of p53 and subsequent cell cycle arrest in G1 and apoptosis in response to oxidative stress (57). Upon depletion of both Artemis and DNA-PKcs, there was a suppression of the phenotype observed upon knockdown of Artemis alone, indicating that Artemis is an inhibitor of DNA-PKcs-mediated stabilization of p53. Artemis cooperates with p53 to suppress tumor formation in multiple tissues (56;58;59). DCLRE1C-deficient pro-B cells that are also deficient in p53 are predisposed to pro-B lymphomas due to oncogenic translocations that are mediated by aberrant V(D)J recombination (58;59). In addition, deficiency in Artemis as well as cell cycle checkpoint defects may result in oncogenic IgH locus translocations during attempted CSR, resulting in B cell lymphomas or multiple myeloma (60). In addition, Dclre1c-deficient Trp53 heterozygous mice develop B lineage lymphomas, osteosarcomas, anaplastic sarcomas and a range of non-malignant pathologies (58).
Mutations in DCLRE1C are linked to severe combined (T−B−NK+) immunodeficiency associated with increased radiosensitivity (RS-SCID; OMIM: #602450), Athabascan SCID (SCID-A; OMIM:#602450), and Omenn syndrome (OS; OMIM: #603554) (1;20;61). Patients with RS-SCID exhibit defects in V(D)J recombination resulting in early maturation defects in B and T cells (1). Patients have absence of complete V(H)-J(H) gene rearrangements and subsequent differentiation arrest of B cells at the pre-BCR checkpoint (61). As a result, the patients display a complete absence of T- and B lymphocytes (62). Some RS-SCID patients have a predisposition to B cell lymphoma (63). Most patients with RS-SCID exhibit early lethality (at approximately 1 year of age) due to opportunistic infections. SCID-A is an autosomal recessive disorder in peoples of the Athabascan-speaking Native Americans (1;20). Similar to RS-SCID, patients present with an absence of both T and B cells due to defective coding joint and precise, but reduced signal joint formation during V(D)J recombination (20). OS is an autosomal recessive condition in which patients present with symptoms of SCID as well as erythrodermia, hepatosplenomegaly, lymphadenopathy, and alopecia (64). OS patients are classified as T+B-NK+ SCID (64). Patients with OS often exhibit elevated or normal T cell counts that are activated and skewed toward a Th2 phenotype (65;66). V(D)J coding joints are normal in the T cells of OS patients. B cells in the OS patients are not detected. The eosinophilia and high IgE levels are the result of increased secretion of the Th2-type cytokines (67). The other immunoglobins were reduced or not detectable in the serum of OS patients. NK cell functions and numbers were unaffected in patients with OS. Patients with OS exhibit lethality; bone marrow transplantation is often successful in treating patients with OS (67).
A DCLRE1C truncation mutation, D451fsX10, results in loss of the C-terminus of Artemis and has been linked to partial immunodeficiency and aggressive EBV-associated lymphoma (63). Patients have low levels of T and B cells, but exhibited lymphocytopenia and died of recurrent infections or lymphoma progression (63).
A spontaneous Dclre1c mutant (68) and Artemis knockout (Dclre1c-/-) mice exhibit early T- and B-cell maturation arrest as well as increased sensitivity to IR (62;69;70). The number of CD11c+MHCII+ dendritic cells, CD3-NK1.1+ NK cells, CD11b+ monocytes and Gr1+ granulocytes were comparable to those in wild-type mice (44;68). In the bone marrow of Dclre1c-/- mice, B cell development was blocked at the B220+/CD43+ progenitor stage resulting in loss of B220+ CD43− precursor and B220+ IgM+ immature B cell (12;62;68;69). Dclre1c-/- mice also lacked peripheral B220+/IgM+ B cells and T cells (44;69;70). Thymocyte numbers were reduced by approximately 50-fold in the Dclre1c-/- mice compared to wild-type or heterozygous littermates. Thymocytes in the Dclre1c-/- mice were predominantly DN T cells, but low numbers of DP and SP thymocytes was observed in some Dclre1c-/- thymocytes indicating that some T cell development occurred (62;68-70). Most thymocytes were arrested in the CD44+CD25- ("DN3") stage of development and few matured to the DP stage; the number of DN1 cells in the thymus were also reduced compared to wild-type levels (44;68). The thymus in the Dclre1c-/- mice did not have a lymphocytic cortex and scattered lymphoid cells with abundant mitotic figures (44). T and B cell frequency were reduced in the lymph nodes and spleens of the Dclre1c-/- mice (44). B cell development was arrested at the early progenitor stage (B220+CD43+) in the lymph nodes and spleens of the Dclre1c-/- mice (44). In the lymph nodes and spleen of some Dclre1cN/N mice, SP T cells (mostly CD4+ CD8−) were observed that also expressed surface CD3 and TCRα/β (69). The spleens from the Dclre1c-/- mice had reduced fully developed lymphoid follicles; most lymphoid cells had larger nuclei with less dense chromatin and prominent nucleoli (44). The lymph nodes from the Dclre1c-/- mice were smaller than those in wild-type mice and were depleted of mature lymphocytes and lymphoid follicles (44). Both T and B cell proliferation were reduced in response to Con A and LPS, respectively in the Dclre1c-/- mice compared to wild-type mice (44).
A targeted Dclre1c mutation (ArtP70/P70; Dclre1ctm1Jsek) in exon 14 resulted in a coding of a premature stop codon at amino acid 449 (Asp449X) and mimics a human DCLRE1C allele, Artemis-P70 (D451X) (12). The ArtemisP70 protein can interact with DNA-PKcs, and it retains exo- and endonuclease activities, but it is not phosphorylated (12). The number of thymocytes and splenocytes in the ArtP70/P70 mice were reduced compared to wild-type and heterozygous mice, but higher than those in Dclre1c knockout (Dclre1c-/-; Dclre1ctm1Fwa) mice (12). The ArtP70/P70 mice exhibited impaired V(D)J recombination, DSB repair, and increased chromosomal instability (12). T cell development was impaired at the DN3 stage, but some T cells did progress to the DP and SP stages (12). In the ArtP70/P70 mice, B cell development was defective at the transition from the pro-B to pre-B stage resulting in reduced percentage and number of pre-B cells than wild-type mice (12). The number of surface IgM-expressing cells in the bone marrow and peripheral lymphoid organs was reduced in the ArtP70/P70 mice (12). The ArtP70/P70 mice displayed a reduced frequency of both D to J and V to DJ rearrangements within the TCR-β locus (12). A modest decrease in the levels of DH to JH rearrangements in the pro- and pre-B cells from ArtP70/P70 mice; significant levels of DH-JH rearrangements occurred (12). Mouse embryonic fibroblasts from the ArtP70/P70 mice exhibited an intermediate hypersensitivity to IR compared to Dclre1c-/- mice (12).
Conditional knockout of Dclre1c in mature B cells (Dclre1ctm2.1Jpdv) resulted in defective switching to certain isotypes (IgG3 and IgA) after B cell activation or after KLH immunization; CSR was not significantly affected (70).
The kiwis mouse phenocopies the previously characterized Dclre1c mutant and knockout models, indicating that the H115R mutation results in loss of Artemis function.
Kiwis genotyping is performed by amplifying the region containing the mutation using PCR, followed by sequencing of the amplified region to detect the single nucleotide transition.
Kiwis_PCR_F: 5’- TACGGGAGTAGACAAGGGTTCCAC -3’
Kiwis_PCR_R: 5’- AAGTAATGCCACCTTTCCCCATGAG -3’
Kiwis_SEQ_F: 5’- GAGTAGACAAGGGTTCCACTTTCTC -3’
Kiwis_SEQ_R: 5’- ACTCAGATCATTGCATTCCAGGG -3’
1) 94°C 2:00
2) 94°C 0:30
3) 55°C 0:30
4) 72°C 1:00
5) repeat steps (2-4) 40X
6) 72°C 10:00
7) 4°C hold
The following sequence of 593 nucleotides is amplified (Genbank genomic region NC_000068):
12483 tacgggag tagacaaggg ttccactttc tctgggtttg gatcatgagg gagtaagaaa
12541 gaggtatgtt cttgtgatac gttgtgaatt tagccacttc ctaatgataa cttccattcc
12601 tgttttgttt tgtttttttg ttttttgttt tttgttttct ttttagaagg aagaggttgt
12661 tgtgactctc ttaccagctg gtcactgccc aggatcagtt atgtaagggg gcccatctgt
12721 ttttgtttct ttctatatat atatatatat atacatatca tatttgtaga aatagctttt
12781 taggatttaa aggtattata tgaacagaaa tagatactct ttttctggca gtgcctttaa
12841 tctttatttc aaaattgagc tcaggaaact gagatgaagg ccatatggag tgtggatgcc
12901 tttaattgat atttcccctg tgactttggc ctgtcttctc atgggcatgt ctctctccct
12961 ggaatgcaat gatctgagtg tatgtatgaa gtgttagact attacaggct acacctagtg
13021 ctcgcccacc ctgccattta ttgtatttat ctcatgggga aaggtggcat tactt
Primer binding sites are underlined and the sequencing primer is highlighted; the mutated nucleotide (A) is shown in red text (A>G).
1. Moshous, D., Callebaut, I., de Chasseval, R., Corneo, B., Cavazzana-Calvo, M., Le Deist, F., Tezcan, I., Sanal, O., Bertrand, Y., Philippe, N., Fischer, A., and de Villartay, J. P. (2001) Artemis, a Novel DNA Double-Strand Break repair/V(D)J Recombination Protein, is Mutated in Human Severe Combined Immune Deficiency. Cell. 105, 177-186.
2. Callebaut, I., Moshous, D., Mornon, J. P., and de Villartay, J. P. (2002) Metallo-Beta-Lactamase Fold within Nucleic Acids Processing Enzymes: The Beta-CASP Family. Nucleic Acids Res. 30, 3592-3601.
3. Kurosawa, A., and Adachi, N. (2010) Functions and Regulation of Artemis: A Goddess in the Maintenance of Genome Integrity. J Radiat Res. 51, 503-509.
4. Yan, Y., Akhter, S., Zhang, X., and Legerski, R. (2010) The Multifunctional SNM1 Gene Family: Not just Nucleases. Future Oncol. 6, 1015-1029.
5. Poinsignon, C., Moshous, D., Callebaut, I., de Chasseval, R., Villey, I., and de Villartay, J. P. (2004) The Metallo-Beta-lactamase/beta-CASP Domain of Artemis Constitutes the Catalytic Core for V(D)J Recombination. J Exp Med. 199, 315-321.
6. Goodarzi, A. A., Yu, Y., Riballo, E., Douglas, P., Walker, S. A., Ye, R., Harer, C., Marchetti, C., Morrice, N., Jeggo, P. A., and Lees-Miller, S. P. (2006) DNA-PK Autophosphorylation Facilitates Artemis Endonuclease Activity. EMBO J. 25, 3880-3889.
7. Ma, Y., Pannicke, U., Lu, H., Niewolik, D., Schwarz, K., and Lieber, M. R. (2005) The DNA-Dependent Protein Kinase Catalytic Subunit Phosphorylation Sites in Human Artemis. J Biol Chem. 280, 33839-33846.
8. Aravind, L. (1999) An Evolutionary Classification of the Metallo-Beta-Lactamase Fold Proteins. In Silico Biol. 1, 69-91.
9. Wang, Z., Fast, W., Valentine, A. M., and Benkovic, S. J. (1999) Metallo-Beta-Lactamase: Structure and Mechanism. Curr Opin Chem Biol. 3, 614-622.
10. Pannicke, U., Ma, Y., Hopfner, K. P., Niewolik, D., Lieber, M. R., and Schwarz, K. (2004) Functional and Biochemical Dissection of the Structure-Specific Nuclease ARTEMIS. EMBO J. 23, 1987-1997.
11. de Villartay, J. P., Shimazaki, N., Charbonnier, J. B., Fischer, A., Mornon, J. P., Lieber, M. R., and Callebaut, I. (2009) A Histidine in the Beta-CASP Domain of Artemis is Critical for its Full in Vitro and in Vivo Functions. DNA Repair (Amst). 8, 202-208.
12. Huang, Y., Giblin, W., Kubec, M., Westfield, G., St Charles, J., Chadde, L., Kraftson, S., and Sekiguchi, J. (2009) Impact of a Hypomorphic Artemis Disease Allele on Lymphocyte Development, DNA End Processing, and Genome Stability. J Exp Med. 206, 893-908.
13. Jacobs, C., Huang, Y., Masud, T., Lu, W., Westfield, G., Giblin, W., and Sekiguchi, J. M. (2011) A Hypomorphic Artemis Human Disease Allele Causes Aberrant Chromosomal Rearrangements and Tumorigenesis. Hum Mol Genet. 20, 806-819.
14. Malu, S., De Ioannes, P., Kozlov, M., Greene, M., Francis, D., Hanna, M., Pena, J., Escalante, C. R., Kurosawa, A., Erdjument-Bromage, H., Tempst, P., Adachi, N., Vezzoni, P., Villa, A., Aggarwal, A. K., and Cortes, P. (2012) Artemis C-Terminal Region Facilitates V(D)J Recombination through its Interactions with DNA Ligase IV and DNA-PKcs. J Exp Med. 209, 955-963.
15. Chen, L., Morio, T., Minegishi, Y., Nakada, S., Nagasawa, M., Komatsu, K., Chessa, L., Villa, A., Lecis, D., Delia, D., and Mizutani, S. (2005) Ataxia-Telangiectasia-Mutated Dependent Phosphorylation of Artemis in Response to DNA Damage. Cancer Sci. 96, 134-141.
16. Zhang, X., Succi, J., Feng, Z., Prithivirajsingh, S., Story, M. D., and Legerski, R. J. (2004) Artemis is a Phosphorylation Target of ATM and ATR and is Involved in the G2/M DNA Damage Checkpoint Response. Mol Cell Biol. 24, 9207-9220.
17. Poinsignon, C., de Chasseval, R., Soubeyrand, S., Moshous, D., Fischer, A., Hache, R. J., and de Villartay, J. P. (2004) Phosphorylation of Artemis Following Irradiation-Induced DNA Damage. Eur J Immunol. 34, 3146-3155.
18. Geng, L., Zhang, X., Zheng, S., and Legerski, R. J. (2007) Artemis Links ATM to G2/M Checkpoint Recovery Via Regulation of Cdk1-Cyclin B. Mol Cell Biol. 27, 2625-2635.
19. Soubeyrand, S., Pope, L., De Chasseval, R., Gosselin, D., Dong, F., de Villartay, J. P., and Hache, R. J. (2006) Artemis Phosphorylated by DNA-Dependent Protein Kinase Associates Preferentially with Discrete Regions of Chromatin. J Mol Biol. 358, 1200-1211.
20. Li, L., Moshous, D., Zhou, Y., Wang, J., Xie, G., Salido, E., Hu, D., de Villartay, J. P., and Cowan, M. J. (2002) A Founder Mutation in Artemis, an SNM1-Like Protein, Causes SCID in Athabascan-Speaking Native Americans. J Immunol. 168, 6323-6329.
21. Mitra, A. K., Bhat, N., Sarma, A., and Krishna, M. (2005) Alteration in the Expression of Signaling Parameters Following Carbon Ion Irradiation. Mol Cell Biochem. 276, 169-173.
22. Msiska, Z., Pacurari, M., Mishra, A., Leonard, S. S., Castranova, V., and Vallyathan, V. (2010) DNA Double-Strand Breaks by Asbestos, Silica, and Titanium Dioxide: Possible Biomarker of Carcinogenic Potential? Am J Respir Cell Mol Biol. 43, 210-219.
23. Saleh-Gohari, N., Bryant, H. E., Schultz, N., Parker, K. M., Cassel, T. N., and Helleday, T. (2005) Spontaneous Homologous Recombination is Induced by Collapsed Replication Forks that are Caused by Endogenous DNA Single-Strand Breaks. Mol Cell Biol. 25, 7158-7169.
24. Meek, K., Gupta, S., Ramsden, D. A., and Lees-Miller, S. P. (2004) The DNA-Dependent Protein Kinase: The Director at the End. Immunol Rev. 200, 132-141.
25. Cahill, D., Connor, B., and Carney, J. P. (2006) Mechanisms of Eukaryotic DNA Double Strand Break Repair. Front Biosci. 11, 1958-1976.
26. Collis, S. J., DeWeese, T. L., Jeggo, P. A., and Parker, A. R. (2005) The Life and Death of DNA-PK. Oncogene. 24, 949-961.
27. Shrivastav, M., De Haro, L. P., and Nickoloff, J. A. (2008) Regulation of DNA Double-Strand Break Repair Pathway Choice. Cell Res. 18, 134-147.
28. Hammarsten, O., DeFazio, L. G., and Chu, G. (2000) Activation of DNA-Dependent Protein Kinase by Single-Stranded DNA Ends. J Biol Chem. 275, 1541-1550.
29. Kaina, B. (2003) DNA Damage-Triggered Apoptosis: Critical Role of DNA Repair, Double-Strand Breaks, Cell Proliferation and Signaling. Biochem Pharmacol. 66, 1547-1554.
30. Elliott, B., and Jasin, M. (2002) Double-Strand Breaks and Translocations in Cancer. Cell Mol Life Sci. 59, 373-385.
31. Khanna, K. K., and Jackson, S. P. (2001) DNA Double-Strand Breaks: Signaling, Repair and the Cancer Connection. Nat Genet. 27, 247-254.
32. Frankenberg-Schwager, M., Gebauer, A., Koppe, C., Wolf, H., Pralle, E., and Frankenberg, D. (2009) Single-Strand Annealing, Conservative Homologous Recombination, Nonhomologous DNA End Joining, and the Cell Cycle-Dependent Repair of DNA Double-Strand Breaks Induced by Sparsely Or Densely Ionizing Radiation. Radiat Res. 171, 265-273.
33. Dip, R., and Naegeli, H. (2005) More than just Strand Breaks: The Recognition of Structural DNA Discontinuities by DNA-Dependent Protein Kinase Catalytic Subunit. FASEB J. 19, 704-715.
34. Rothkamm, K., Kruger, I., Thompson, L. H., and Lobrich, M. (2003) Pathways of DNA Double-Strand Break Repair during the Mammalian Cell Cycle. Mol Cell Biol. 23, 5706-5715.
35. Beucher, A., Birraux, J., Tchouandong, L., Barton, O., Shibata, A., Conrad, S., Goodarzi, A. A., Krempler, A., Jeggo, P. A., and Lobrich, M. (2009) ATM and Artemis Promote Homologous Recombination of Radiation-Induced DNA Double-Strand Breaks in G2. EMBO J. 28, 3413-3427.
36. Lieber, M. R. (2008) The Mechanism of Human Nonhomologous DNA End Joining. J Biol Chem. 283, 1-5.
37. Weterings, E., and Chen, D. J. (2007) DNA-Dependent Protein Kinase in Nonhomologous End Joining: A Lock with Multiple Keys? J Cell Biol. 179, 183-186.
38. Malyarchuk, S., Castore, R., Shi, R., and Harrison, L. (2013) Artemis is Required to Improve the Accuracy of Repair of Double-Strand Breaks with 5'-Blocked Termini Generated from Non-DSB-Clustered Lesions. Mutagenesis. 28, 357-366.
39. Ma, Y., Pannicke, U., Schwarz, K., and Lieber, M. R. (2002) Hairpin Opening and Overhang Processing by an Artemis/DNA-Dependent Protein Kinase Complex in Nonhomologous End Joining and V(D)J Recombination. Cell. 108, 781-794.
40. Povirk, L. F., Zhou, T., Zhou, R., Cowan, M. J., and Yannone, S. M. (2007) Processing of 3'-Phosphoglycolate-Terminated DNA Double Strand Breaks by Artemis Nuclease. J Biol Chem. 282, 3547-3558.
41. Weterings, E., Verkaik, N. S., Keijzers, G., Florea, B. I., Wang, S. Y., Ortega, L. G., Uematsu, N., Chen, D. J., and van Gent, D. C. (2009) The Ku80 Carboxy Terminus Stimulates Joining and Artemis-Mediated Processing of DNA Ends. Mol Cell Biol. 29, 1134-1142.
42. Yannone, S. M., Khan, I. S., Zhou, R. Z., Zhou, T., Valerie, K., and Povirk, L. F. (2008) Coordinate 5' and 3' Endonucleolytic Trimming of Terminally Blocked Blunt DNA Double-Strand Break Ends by Artemis Nuclease and DNA-Dependent Protein Kinase. Nucleic Acids Res. 36, 3354-3365.
43. Niewolik, D., Pannicke, U., Lu, H., Ma, Y., Wang, L. C., Kulesza, P., Zandi, E., Lieber, M. R., and Schwarz, K. (2006) DNA-PKcs Dependence of Artemis Endonucleolytic Activity, Differences between Hairpins and 5' Or 3' Overhangs. J Biol Chem. 281, 33900-33909.
44. Li, L., Salido, E., Zhou, Y., Bhattacharyya, S., Yannone, S. M., Dunn, E., Meneses, J., Feeney, A. J., and Cowan, M. J. (2005) Targeted Disruption of the Artemis Murine Counterpart Results in SCID and Defective V(D)J Recombination that is Partially Corrected with Bone Marrow Transplantation. J Immunol. 174, 2420-2428.
45. Rooney, S., Alt, F. W., Sekiguchi, J., and Manis, J. P. (2005) Artemis-Independent Functions of DNA-Dependent Protein Kinase in Ig Heavy Chain Class Switch Recombination and Development. Proc Natl Acad Sci U S A. 102, 2471-2475.
46. Rooney, S., Alt, F. W., Lombard, D., Whitlow, S., Eckersdorff, M., Fleming, J., Fugmann, S., Ferguson, D. O., Schatz, D. G., and Sekiguchi, J. (2003) Defective DNA Repair and Increased Genomic Instability in Artemis-Deficient Murine Cells. J Exp Med. 197, 553-565.
47. Manis, J. P., Morales, J. C., Xia, Z., Kutok, J. L., Alt, F. W., and Carpenter, P. B. (2004) 53BP1 Links DNA Damage-Response Pathways to Immunoglobulin Heavy Chain Class-Switch Recombination. Nat Immunol. 5, 481-487.
48. Ward, I. M., Reina-San-Martin, B., Olaru, A., Minn, K., Tamada, K., Lau, J. S., Cascalho, M., Chen, L., Nussenzweig, A., Livak, F., Nussenzweig, M. C., and Chen, J. (2004) 53BP1 is Required for Class Switch Recombination. J Cell Biol. 165, 459-464.
49. Stavnezer, J., Guikema, J. E., and Schrader, C. E. (2008) Mechanism and Regulation of Class Switch Recombination. Annu Rev Immunol. 26, 261-292.
50. Bothmer, A., Robbiani, D. F., Feldhahn, N., Gazumyan, A., Nussenzweig, A., and Nussenzweig, M. C. (2010) 53BP1 Regulates DNA Resection and the Choice between Classical and Alternative End Joining during Class Switch Recombination. J Exp Med. 207, 855-865.
51. Bothmer, A., Rommel, P. C., Gazumyan, A., Polato, F., Reczek, C. R., Muellenbeck, M. F., Schaetzlein, S., Edelmann, W., Chen, P. L., Brosh, R. M.,Jr, Casellas, R., Ludwig, T., Baer, R., Nussenzweig, A., Nussenzweig, M. C., and Robbiani, D. F. (2013) Mechanism of DNA Resection during Intrachromosomal Recombination and Immunoglobulin Class Switching. J Exp Med. 210, 115-123.
52. Franco, S., Murphy, M. M., Li, G., Borjeson, T., Boboila, C., and Alt, F. W. (2008) DNA-PKcs and Artemis Function in the End-Joining Phase of Immunoglobulin Heavy Chain Class Switch Recombination. J Exp Med. 205, 557-564.
53. Du, L., van der Burg, M., Popov, S. W., Kotnis, A., van Dongen, J. J., Gennery, A. R., and Pan-Hammarstrom, Q. (2008) Involvement of Artemis in Nonhomologous End-Joining during Immunoglobulin Class Switch Recombination. J Exp Med. 205, 3031-3040.
54. Wang, H., Zhang, X., Geng, L., Teng, L., and Legerski, R. J. (2009) Artemis Regulates Cell Cycle Recovery from the S Phase Checkpoint by Promoting Degradation of Cyclin E. J Biol Chem. 284, 18236-18243.
55. Yan, Y., Zhang, X., and Legerski, R. J. (2011) Artemis Interacts with the Cul4A-DDB1DDB2 Ubiquitin E3 Ligase and Regulates Degradation of the CDK Inhibitor p27. Cell Cycle. 10, 4098-4109.
56. Maas, S. A., Donghia, N. M., Tompkins, K., Foreman, O., and Mills, K. D. (2010) ARTEMIS Stabilizes the Genome and Modulates Proliferative Responses in Multipotent Mesenchymal Cells. BMC Biol. 8, 132-7007-8-132.
57. Zhang, X., Zhu, Y., Geng, L., Wang, H., and Legerski, R. J. (2009) Artemis is a Negative Regulator of p53 in Response to Oxidative Stress. Oncogene. 28, 2196-2204.
58. Woo, Y., Wright, S. M., Maas, S. A., Alley, T. L., Caddle, L. B., Kamdar, S., Affourtit, J., Foreman, O., Akeson, E. C., Shaffer, D., Bronson, R. T., Morse, H. C.,3rd, Roopenian, D., and Mills, K. D. (2007) The Nonhomologous End Joining Factor Artemis Suppresses Multi-Tissue Tumor Formation and Prevents Loss of Heterozygosity. Oncogene. 26, 6010-6020.
59. Rooney, S., Sekiguchi, J., Whitlow, S., Eckersdorff, M., Manis, J. P., Lee, C., Ferguson, D. O., and Alt, F. W. (2004) Artemis and p53 Cooperate to Suppress Oncogenic N-Myc Amplification in Progenitor B Cells. Proc Natl Acad Sci U S A. 101, 2410-2415.
60. Ramiro, A. R., Jankovic, M., Callen, E., Difilippantonio, S., Chen, H. T., McBride, K. M., Eisenreich, T. R., Chen, J., Dickins, R. A., Lowe, S. W., Nussenzweig, A., and Nussenzweig, M. C. (2006) Role of Genomic Instability and p53 in AID-Induced c-Myc-Igh Translocations. Nature. 440, 105-109.
61. Noordzij, J. G., Verkaik, N. S., van der Burg, M., van Veelen, L. R., de Bruin-Versteeg, S., Wiegant, W., Vossen, J. M., Weemaes, C. M., de Groot, R., Zdzienicka, M. Z., van Gent, D. C., and van Dongen, J. J. (2003) Radiosensitive SCID Patients with Artemis Gene Mutations show a Complete B-Cell Differentiation Arrest at the Pre-B-Cell Receptor Checkpoint in Bone Marrow. Blood. 101, 1446-1452.
62. Benjelloun, F., Garrigue, A., Demerens-de Chappedelaine, C., Soulas-Sprauel, P., Malassis-Seris, M., Stockholm, D., Hauer, J., Blondeau, J., Riviere, J., Lim, A., Le Lorc'h, M., Romana, S., Brousse, N., Paques, F., Galy, A., Charneau, P., Fischer, A., de Villartay, J. P., and Cavazzana-Calvo, M. (2008) Stable and Functional Lymphoid Reconstitution in Artemis-Deficient Mice Following Lentiviral Artemis Gene Transfer into Hematopoietic Stem Cells. Mol Ther. 16, 1490-1499.
63. Moshous, D., Pannetier, C., Chasseval Rd, R., Deist Fl, F., Cavazzana-Calvo, M., Romana, S., Macintyre, E., Canioni, D., Brousse, N., Fischer, A., Casanova, J. L., and Villartay, J. P. (2003) Partial T and B Lymphocyte Immunodeficiency and Predisposition to Lymphoma in Patients with Hypomorphic Mutations in Artemis. J Clin Invest. 111, 381-387.
64. Ege, M., Ma, Y., Manfras, B., Kalwak, K., Lu, H., Lieber, M. R., Schwarz, K., and Pannicke, U. (2005) Omenn Syndrome due to ARTEMIS Mutations. Blood. 105, 4179-4186.
65. Schandene, L., Ferster, A., Mascart-Lemone, F., Crusiaux, A., Gerard, C., Marchant, A., Lybin, M., Velu, T., Sariban, E., and Goldman, M. (1993) T Helper Type 2-Like Cells and Therapeutic Effects of Interferon-Gamma in Combined Immunodeficiency with Hypereosinophilia (Omenn's Syndrome). Eur J Immunol. 23, 56-60.
66. Chilosi, M., Facchetti, F., Notarangelo, L. D., Romagnani, S., Del Prete, G., Almerigogna, F., De Carli, M., and Pizzolo, G. (1996) CD30 Cell Expression and Abnormal Soluble CD30 Serum Accumulation in Omenn's Syndrome: Evidence for a T Helper 2-Mediated Condition. Eur J Immunol. 26, 329-334.
67. Aleman, K., Noordzij, J. G., de Groot, R., van Dongen, J. J., and Hartwig, N. G. (2001) Reviewing Omenn Syndrome. Eur J Pediatr. 160, 718-725.
68. Barthels, C., Puchalka, J., Racek, T., Klein, C., and Brocker, T. (2013) Novel Spontaneous Deletion of Artemis Exons 10 and 11 in Mice Leads to T- and B-Cell Deficiency. PLoS One. 8, e74838.
69. Rooney, S., Sekiguchi, J., Zhu, C., Cheng, H. L., Manis, J., Whitlow, S., DeVido, J., Foy, D., Chaudhuri, J., Lombard, D., and Alt, F. W. (2002) Leaky Scid Phenotype Associated with Defective V(D)J Coding End Processing in Artemis-Deficient Mice. Mol Cell. 10, 1379-1390.
|Science Writers||Anne Murray|
|Authors||Kuan-Wen Wang, Jin Huk Choi, Ming Zeng, Bruce Beutler|