Phenotypic Mutation 'abra2' (pdf version)
Allele | abra2 |
Mutation Type |
splice site
|
Chromosome | 2 |
Coordinate | 126,762,233 bp (GRCm39) |
Base Change | C ⇒ T (forward strand) |
Gene |
Sppl2a
|
Gene Name | signal peptide peptidase like 2A |
Synonym(s) | C130089K23Rik, 2010106G01Rik |
Chromosomal Location |
126,732,311-126,775,155 bp (-) (GRCm39)
|
MGI Phenotype |
FUNCTION: [Summary is not available for the mouse gene. This summary is for the human ortholog.] This gene encodes a member of the GXGD family of aspartic proteases, which are transmembrane proteins with two conserved catalytic motifs localized within the membrane-spanning regions, as well as a member of the signal peptide peptidase-like protease (SPPL) family. This protein is expressed in all major adult human tissues and localizes to late endosomal compartments and lysosomal membranes. A pseudogene of this gene also lies on chromosome 15. [provided by RefSeq, Feb 2012] PHENOTYPE: Mice homozygous for a knock-out allele exhibit decreased immunoglobulin prior to and after immunization and decreased splenic B cells, myeloid dendritic cells, T2 B cells and follicular B cells. Mice homozygous for a hypomorphic allele exhibit similar albeit less severe phenotypes. [provided by MGI curators]
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Accession Number | NCBI RefSeq: NM_023220; MGI:1913802
|
Mapped | Yes |
Limits of the Critical Region |
126890391 - 126933235 bp |
Amino Acid Change |
|
Institutional Source | Beutler Lab |
Gene Model |
predicted gene model for protein(s):
[ENSMUSP00000028844 †]
† probably from a misspliced transcript
|
AlphaFold |
Q9JJF9 |
SMART Domains |
Protein: ENSMUSP00000028844 Gene: ENSMUSG00000027366
Domain | Start | End | E-Value | Type |
signal peptide
|
1 |
25 |
N/A |
INTRINSIC |
Pfam:PA
|
58 |
153 |
1.7e-12 |
PFAM |
transmembrane domain
|
173 |
195 |
N/A |
INTRINSIC |
PSN
|
218 |
486 |
3.65e-102 |
SMART |
|
Predicted Effect |
probably null
|
SMART Domains |
Protein: ENSMUSP00000119064 Gene: ENSMUSG00000027366
Domain | Start | End | E-Value | Type |
PSN
|
3 |
233 |
1.27e-60 |
SMART |
|
Predicted Effect |
probably null
|
Meta Mutation Damage Score |
0.9755 |
Is this an essential gene? |
Probably nonessential (E-score: 0.080) |
Phenotypic Category |
Autosomal Recessive |
Candidate Explorer Status |
loading ... |
Single pedigree Linkage Analysis Data
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|
Penetrance | |
Alleles Listed at MGI | All Mutations and Alleles(14) : Chemically induced (ENU)(2) Chemically induced (other)(1) Gene trapped(6) Radiation induced(1) Targeted(4)
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Lab Alleles |
Allele | Source | Chr | Coord | Type | Predicted Effect | PPH Score |
IGL00781:Sppl2a
|
APN |
2 |
126761640 |
missense |
probably benign |
0.04 |
IGL01471:Sppl2a
|
APN |
2 |
126759787 |
nonsense |
probably null |
|
IGL01572:Sppl2a
|
APN |
2 |
126762232 |
splice site |
probably null |
|
IGL01712:Sppl2a
|
APN |
2 |
126746823 |
splice site |
probably benign |
|
IGL02203:Sppl2a
|
APN |
2 |
126746861 |
missense |
possibly damaging |
0.68 |
IGL02572:Sppl2a
|
APN |
2 |
126768216 |
missense |
probably benign |
0.07 |
abra
|
UTSW |
2 |
126765514 |
missense |
probably benign |
0.00 |
isaac
|
UTSW |
2 |
126755495 |
missense |
probably damaging |
1.00 |
jacob
|
UTSW |
2 |
126755201 |
splice site |
probably null |
|
PIT4431001:Sppl2a
|
UTSW |
2 |
126765396 |
missense |
probably damaging |
1.00 |
R0023:Sppl2a
|
UTSW |
2 |
126755213 |
splice site |
probably null |
|
R0240:Sppl2a
|
UTSW |
2 |
126762256 |
missense |
probably benign |
0.14 |
R0240:Sppl2a
|
UTSW |
2 |
126762256 |
missense |
probably benign |
0.14 |
R0458:Sppl2a
|
UTSW |
2 |
126746879 |
missense |
probably damaging |
1.00 |
R0627:Sppl2a
|
UTSW |
2 |
126762337 |
unclassified |
probably benign |
|
R0799:Sppl2a
|
UTSW |
2 |
126762227 |
splice site |
probably benign |
|
R1029:Sppl2a
|
UTSW |
2 |
126765514 |
missense |
probably benign |
0.00 |
R1245:Sppl2a
|
UTSW |
2 |
126755441 |
splice site |
probably benign |
|
R1669:Sppl2a
|
UTSW |
2 |
126759714 |
splice site |
probably benign |
|
R2047:Sppl2a
|
UTSW |
2 |
126768772 |
missense |
probably damaging |
1.00 |
R2215:Sppl2a
|
UTSW |
2 |
126769754 |
missense |
probably benign |
0.00 |
R2428:Sppl2a
|
UTSW |
2 |
126754615 |
missense |
possibly damaging |
0.93 |
R3522:Sppl2a
|
UTSW |
2 |
126762242 |
missense |
possibly damaging |
0.66 |
R4653:Sppl2a
|
UTSW |
2 |
126762233 |
splice site |
probably null |
|
R5398:Sppl2a
|
UTSW |
2 |
126761638 |
missense |
probably benign |
0.00 |
R6382:Sppl2a
|
UTSW |
2 |
126758949 |
splice site |
probably null |
|
R6888:Sppl2a
|
UTSW |
2 |
126746912 |
missense |
probably damaging |
0.99 |
R6892:Sppl2a
|
UTSW |
2 |
126755495 |
missense |
probably damaging |
1.00 |
R7021:Sppl2a
|
UTSW |
2 |
126769663 |
splice site |
probably null |
|
R7750:Sppl2a
|
UTSW |
2 |
126761625 |
missense |
probably damaging |
1.00 |
R8129:Sppl2a
|
UTSW |
2 |
126765390 |
missense |
probably damaging |
1.00 |
R8136:Sppl2a
|
UTSW |
2 |
126755201 |
splice site |
probably null |
|
R8772:Sppl2a
|
UTSW |
2 |
126768231 |
missense |
probably benign |
0.16 |
R9128:Sppl2a
|
UTSW |
2 |
126765393 |
missense |
probably damaging |
1.00 |
R9144:Sppl2a
|
UTSW |
2 |
126769743 |
missense |
probably benign |
0.00 |
RF016:Sppl2a
|
UTSW |
2 |
126769694 |
missense |
probably benign |
0.01 |
|
Mode of Inheritance |
Autosomal Recessive |
Local Stock | |
Repository | |
Last Updated |
2019-09-04 9:42 PM
by Anne Murray
|
Record Created |
2016-06-23 10:42 PM
by Jin Huk Choi
|
Record Posted |
2016-10-28 |
Phenotypic Description |
The abra2 phenotype was identified among G3 mice of the pedigree R4653, some of which showed diminished T-dependent antibody response to ovalbumin administered with aluminum hydroxide (Figure 1) and to recombinant Semliki Forest virus (rSFV)-encoded β-galactosidase (rSFV-β-gal) (Figure 2) were diminished. The T-independent antibody response to 4-hydroxy-3-nitrophenylacetyl-Ficoll (NP-Ficoll) was also diminished (Figure 3).
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Nature of Mutation |
Whole exome HiSeq sequencing of the G1 grandsire identified 62 mutations. All of the above anomalies were linked by continuous variable mapping to a mutation in three genes on chromosome 2: Ext2, Ryr3, and Sppl2a. The Sppl2a mutation is presumed causative, because the immune phenotypes observed in abra2 mimic those listed on MGI and another Sppl2a mutant identified in a second pedigree (see abra). The Sppl2a mutation is a G to A transition at base pair 126,920,313 (v38) on chromosome 2, or base pair 12,950 in the GenBank genomic region NC_000068 encoding the Sppl2a gene. The strongest association was found with a recessive model of linkage to the normalized antibody response to rSFV-β-gal (P = 7.067 x 10-15), wherein four variant homozygotes departed phenotypically from four homozygous reference mice and four heterozygous mice (Figure 4). The effect of the mutation at the cDNA and protein level have not examined, but the mutation is predicted to result in the use of a cryptic site in intron 7. Use of the cryptic splice site would result in a 54-base pair insertion in intron 7, leading to a frame-shifted protein product beginning after amino acid 281, and termination after the inclusion of 9 aberrant amino acids.
C57BL/6J:
<--exon 6 <--exon 7 intron 7--> exon 8-->
……TTCTACAGGTGGCTTG ……TGTGGACAGTGCAC gtacgtatctgtt…… GATTTTGTGTTGTGG……
244 ……-F--Y--R--W--L-- ……-C--G--Q--C--T --I--L--C--C--G…
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Genomic numbering corresponds to NC_000068. The donor splice site of intron 7, which is destroyed by the abra2 mutation, is indicated in blue lettering and the mutated nucleotide is indicated in red.
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Illustration of Mutations in
Gene & Protein |
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Protein Prediction |
Sppl2a encodes SPPL2a, a signal peptide peptidase-like (SPPL) protease, and member of the GxGD intramembrane-cleaving protease family of aspartyl intramembrane-cleaving proteases (I-CLiPs) (Figure 5). SPPL proteins share a YD motif (amino acids 354-355), a GXGD motif (amino acids 413-416), and a PAL sequence (amino acids 466-468) (1-4). SPPL2A has nine transmembrane domains. The abra2 mutation is within intron 7. For more information about Sppl2a, please see the record for abra.
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Putative Mechanism | GxGD proteases are essential for the regulated intramembrane proteolysis (RIP) of single span type II transmembrane proteins. SPPL2A cleaves TNF-α (see the record for PanR1; (5)), ITM2B (Bri2) (6), Fas ligand (FasL; see the record for riogrande) (7), CD74 (8-13), TMEM106B (14), and neuregulin 1 type III (15). Sppl2a knockout (Sppl2a-/-) mice have a developmental block during the splenic phase of B cell maturation. As a result, the frequency of mature and functionally competent B cells is reduced, and the mice exhibit impaired humoral immune responses (8). Abra2 mice exhibited defects in the T-independent B cell response to NP-Ficoll as well as T-dependent antibody responses to rSFV-β-gal and OVA-alum. Taken together, the abra2 mutation results in loss of SPPL2A function.
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Primers |
PCR Primer
abra2_pcr_F: TCAGGAATTCAGTGTAAAGATCCC
abra2_pcr_R: AGACTCTTGGTTGCGCCTTAG
Sequencing Primer
abra2_seq_F: CCTTTAAGTCCTAGCACTGAGGTAG
abra2_seq_R: TGCGCCTTAGCAACTGC
|
Genotyping | PCR program 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 563 nucleotides is amplified (chromosome 2, - strand):
1 agactcttgg ttgcgcctta gcaactgctt cacatagcaa tatagttact atatagaaag 61 tatgatctaa tgaaaagagc taaggaatta acatctgtct tgttcttttc tagtgtatgt 121 tatgatagcg attttctgca tagcatcgtc aatgagcctg tacaactgtc tttctgcatt 181 gattcatagg atgccatgtg gacagtgcac gtacgtatct gttagcctat gatgcaatta 241 gtatttcagc ttagtacagg aaaagtgagg tgttgcttta agcctgtata ttcaagattg 301 tatttatgcc tgtctgagtt taaagttaat aatgtgtttg ggattttttt gtttgttttg 361 ttttatgaga cagggtttct cgggagccta gttgtcctgg aactgtctct gtagacctgg 421 ctggcctcga actcacagag atctatctgt ctgcctctgc ctcctaagtt caggaacaag 481 gtgtgcacta cctcagtgct aggacttaaa ggtgtgtgcc accatgccca gatgtgtttg 541 ggatctttac actgaattcc tga
Primer binding sites are underlined and the sequencing primers are highlighted; the mutated nucleotide is shown in red. |
References | 1. Weihofen, A., Binns, K., Lemberg, M. K., Ashman, K., and Martoglio, B. (2002) Identification of Signal Peptide Peptidase, a Presenilin-Type Aspartic Protease. Science. 296, 2215-2218.
2. Ponting, C. P., Hutton, M., Nyborg, A., Baker, M., Jansen, K., and Golde, T. E. (2002) Identification of a Novel Family of Presenilin Homologues. Hum Mol Genet. 11, 1037-1044.
5. Friedmann, E., Hauben, E., Maylandt, K., Schleeger, S., Vreugde, S., Lichtenthaler, S. F., Kuhn, P. H., Stauffer, D., Rovelli, G., and Martoglio, B. (2006) SPPL2a and SPPL2b Promote Intramembrane Proteolysis of TNFalpha in Activated Dendritic Cells to Trigger IL-12 Production. Nat Cell Biol. 8, 843-848.
6. Martin, L., Fluhrer, R., Reiss, K., Kremmer, E., Saftig, P., and Haass, C. (2008) Regulated Intramembrane Proteolysis of Bri2 (Itm2b) by ADAM10 and SPPL2a/SPPL2b. J Biol Chem. 283, 1644-1652.
7. Kirkin, V., Cahuzac, N., Guardiola-Serrano, F., Huault, S., Luckerath, K., Friedmann, E., Novac, N., Wels, W. S., Martoglio, B., Hueber, A. O., and Zornig, M. (2007) The Fas Ligand Intracellular Domain is Released by ADAM10 and SPPL2a Cleavage in T-Cells. Cell Death Differ. 14, 1678-1687.
8. Schneppenheim, J., Dressel, R., Huttl, S., Lullmann-Rauch, R., Engelke, M., Dittmann, K., Wienands, J., Eskelinen, E. L., Hermans-Borgmeyer, I., Fluhrer, R., Saftig, P., and Schroder, B. (2013) The Intramembrane Protease SPPL2a Promotes B Cell Development and Controls Endosomal Traffic by Cleavage of the Invariant Chain. J Exp Med. 210, 41-58.
9. Bergmann, H., Yabas, M., Short, A., Miosge, L., Barthel, N., Teh, C. E., Roots, C. M., Bull, K. R., Jeelall, Y., Horikawa, K., Whittle, B., Balakishnan, B., Sjollema, G., Bertram, E. M., Mackay, F., Rimmer, A. J., Cornall, R. J., Field, M. A., Andrews, T. D., Goodnow, C. C., and Enders, A. (2013) B Cell Survival, Surface BCR and BAFFR Expression, CD74 Metabolism, and CD8- Dendritic Cells Require the Intramembrane Endopeptidase SPPL2A. J Exp Med. 210, 31-40.
10. Beisner, D. R., Langerak, P., Parker, A. E., Dahlberg, C., Otero, F. J., Sutton, S. E., Poirot, L., Barnes, W., Young, M. A., Niessen, S., Wiltshire, T., Bodendorf, U., Martoglio, B., Cravatt, B., and Cooke, M. P. (2013) The Intramembrane Protease Sppl2a is Required for B Cell and DC Development and Survival Via Cleavage of the Invariant Chain. J Exp Med. 210, 23-30.
11. Huttl, S., Klasener, K., Schweizer, M., Schneppenheim, J., Oberg, H. H., Kabelitz, D., Reth, M., Saftig, P., and Schroder, B. (2015) Processing of CD74 by the Intramembrane Protease SPPL2a is Critical for B Cell Receptor Signaling in Transitional B Cells. J Immunol. 195, 1548-1563.
12. Schneppenheim, J., Huttl, S., Kruchen, A., Fluhrer, R., Muller, I., Saftig, P., Schneppenheim, R., Martin, C. L., and Schroder, B. (2014) Signal-Peptide-Peptidase-Like 2a is Required for CD74 Intramembrane Proteolysis in Human B Cells. Biochem Biophys Res Commun. 451, 48-53.
13. Oliveira, C. C., Querido, B., Sluijter, M., de Groot, A. F., van der Zee, R., Rabelink, M. J., Hoeben, R. C., Ossendorp, F., van der Burg, S. H., and van Hall, T. (2013) New Role of Signal Peptide Peptidase to Liberate C-Terminal Peptides for MHC Class I Presentation. J Immunol. 191, 4020-4028.
15. Fleck, D., Voss, M., Brankatschk, B., Giudici, C., Hampel, H., Schwenk, B., Edbauer, D., Fukumori, A., Steiner, H., Kremmer, E., Haug-Kroper, M., Rossner, M. J., Fluhrer, R., Willem, M., and Haass, C. (2016) Proteolytic Processing of Neuregulin 1 Type III by Three Intramembrane-Cleaving Proteases. J Biol Chem. 291, 318-333.
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Science Writers | Anne Murray |
Illustrators | Katherine Timer |
Authors | Jin Huk Choi, James Butler, Bruce Beutler |