Phenotypic Mutation 'untersuchen' (pdf version)
Allele | untersuchen |
Mutation Type |
missense
|
Chromosome | 4 |
Coordinate | 65,215,494 bp (GRCm39) |
Base Change | T ⇒ A (forward strand) |
Gene |
Pappa
|
Gene Name | pregnancy-associated plasma protein A |
Synonym(s) | PAPP-A, PAG1, 8430414N03Rik, IGFBP-4ase |
Chromosomal Location |
65,042,411-65,275,746 bp (+) (GRCm39)
|
MGI Phenotype |
FUNCTION: [Summary is not available for the mouse gene. This summary is for the human ortholog.] This gene encodes a secreted metalloproteinase which cleaves insulin-like growth factor binding proteins (IGFBPs). It is thought to be involved in local proliferative processes such as wound healing and bone remodeling. Low plasma level of this protein has been suggested as a biochemical marker for pregnancies with aneuploid fetuses. [provided by RefSeq, Jul 2008] PHENOTYPE: Homozygous null mutants are smaller than normal with delayed ossification, but are otherwise normal and fertile. [provided by MGI curators]
|
Accession Number | NCBI RefSeq: NM_021362; MGI:97479
|
Mapped | Yes |
Amino Acid Change |
Leucine changed to Methionine
|
Institutional Source | Beutler Lab |
Gene Model |
predicted gene model for protein(s):
[ENSMUSP00000081545]
|
AlphaFold |
Q8R4K8 |
SMART Domains |
Protein: ENSMUSP00000081545 Gene: ENSMUSG00000028370 AA Change: L1134M
Domain | Start | End | E-Value | Type |
signal peptide
|
1 |
22 |
N/A |
INTRINSIC |
low complexity region
|
24 |
66 |
N/A |
INTRINSIC |
low complexity region
|
69 |
87 |
N/A |
INTRINSIC |
LamGL
|
114 |
263 |
1.55e-54 |
SMART |
NL
|
396 |
438 |
4.15e-8 |
SMART |
NL
|
441 |
471 |
6.73e-1 |
SMART |
Pfam:Peptidase_M43
|
500 |
657 |
2.5e-10 |
PFAM |
Blast:FN3
|
669 |
929 |
1e-165 |
BLAST |
CCP
|
1212 |
1277 |
1.39e-9 |
SMART |
CCP
|
1282 |
1339 |
1.08e-6 |
SMART |
CCP
|
1343 |
1407 |
1.64e-6 |
SMART |
CCP
|
1412 |
1468 |
8.06e-6 |
SMART |
NL
|
1544 |
1581 |
3.24e-10 |
SMART |
low complexity region
|
1584 |
1591 |
N/A |
INTRINSIC |
|
Predicted Effect |
probably damaging
PolyPhen 2
Score 1.000 (Sensitivity: 0.00; Specificity: 1.00)
(Using ENSMUST00000084501)
|
Meta Mutation Damage Score |
0.6467 |
Is this an essential gene? |
Non Essential (E-score: 0.000) |
Phenotypic Category |
Unknown |
Candidate Explorer Status |
loading ... |
Single pedigree Linkage Analysis Data
|
|
Penetrance | |
Alleles Listed at MGI | All Mutations and Alleles(7) : Chemically induced (ENU)(3) Targeted (4)
|
Lab Alleles |
Allele | Source | Chr | Coord | Type | Predicted Effect | PPH Score |
IGL01096:Pappa
|
APN |
4 |
65107553 |
missense |
probably damaging |
1.00 |
IGL01340:Pappa
|
APN |
4 |
65242109 |
missense |
possibly damaging |
0.49 |
IGL01482:Pappa
|
APN |
4 |
65074271 |
missense |
probably benign |
0.18 |
IGL01485:Pappa
|
APN |
4 |
65107536 |
missense |
probably damaging |
0.96 |
IGL01759:Pappa
|
APN |
4 |
65123395 |
splice site |
probably null |
|
IGL01860:Pappa
|
APN |
4 |
65123329 |
missense |
possibly damaging |
0.50 |
IGL01990:Pappa
|
APN |
4 |
65074924 |
splice site |
probably benign |
|
IGL02089:Pappa
|
APN |
4 |
65074361 |
missense |
possibly damaging |
0.75 |
IGL02153:Pappa
|
APN |
4 |
65215674 |
missense |
probably damaging |
0.96 |
IGL02184:Pappa
|
APN |
4 |
65258928 |
missense |
possibly damaging |
0.82 |
IGL02324:Pappa
|
APN |
4 |
65115045 |
missense |
probably damaging |
0.99 |
IGL02542:Pappa
|
APN |
4 |
65094518 |
missense |
probably damaging |
1.00 |
IGL02556:Pappa
|
APN |
4 |
65074863 |
missense |
possibly damaging |
0.56 |
IGL02698:Pappa
|
APN |
4 |
65099257 |
missense |
probably damaging |
1.00 |
IGL02903:Pappa
|
APN |
4 |
65180217 |
missense |
probably damaging |
1.00 |
IGL02974:Pappa
|
APN |
4 |
65123172 |
missense |
probably damaging |
1.00 |
IGL03107:Pappa
|
APN |
4 |
65122940 |
missense |
probably damaging |
1.00 |
IGL03376:Pappa
|
APN |
4 |
65115071 |
missense |
probably benign |
0.01 |
caer
|
UTSW |
4 |
65043128 |
missense |
probably damaging |
0.98 |
Maennel
|
UTSW |
4 |
65232824 |
missense |
probably benign |
0.05 |
maennelein
|
UTSW |
4 |
65233033 |
splice site |
probably null |
|
mama
|
UTSW |
4 |
65123104 |
missense |
possibly damaging |
0.94 |
Revisitation
|
UTSW |
4 |
65212705 |
missense |
probably damaging |
0.96 |
Sesquester
|
UTSW |
4 |
65074612 |
missense |
possibly damaging |
0.66 |
IGL02980:Pappa
|
UTSW |
4 |
65226011 |
missense |
probably benign |
0.25 |
PIT4498001:Pappa
|
UTSW |
4 |
65234469 |
missense |
probably damaging |
1.00 |
R0077:Pappa
|
UTSW |
4 |
65226049 |
missense |
probably damaging |
1.00 |
R0390:Pappa
|
UTSW |
4 |
65269850 |
splice site |
probably null |
|
R0458:Pappa
|
UTSW |
4 |
65074119 |
missense |
probably damaging |
1.00 |
R0883:Pappa
|
UTSW |
4 |
65107552 |
nonsense |
probably null |
|
R0946:Pappa
|
UTSW |
4 |
65233029 |
critical splice donor site |
probably null |
|
R1228:Pappa
|
UTSW |
4 |
65258926 |
missense |
probably damaging |
1.00 |
R1327:Pappa
|
UTSW |
4 |
65269840 |
splice site |
probably benign |
|
R1489:Pappa
|
UTSW |
4 |
65099185 |
missense |
possibly damaging |
0.85 |
R1619:Pappa
|
UTSW |
4 |
65094466 |
missense |
probably damaging |
1.00 |
R1856:Pappa
|
UTSW |
4 |
65258980 |
missense |
probably damaging |
1.00 |
R2047:Pappa
|
UTSW |
4 |
65149378 |
splice site |
probably benign |
|
R2102:Pappa
|
UTSW |
4 |
65234465 |
nonsense |
probably null |
|
R2127:Pappa
|
UTSW |
4 |
65215494 |
missense |
probably damaging |
1.00 |
R2143:Pappa
|
UTSW |
4 |
65099186 |
nonsense |
probably null |
|
R2144:Pappa
|
UTSW |
4 |
65099186 |
nonsense |
probably null |
|
R2166:Pappa
|
UTSW |
4 |
65074682 |
missense |
probably damaging |
1.00 |
R2167:Pappa
|
UTSW |
4 |
65074682 |
missense |
probably damaging |
1.00 |
R2168:Pappa
|
UTSW |
4 |
65074682 |
missense |
probably damaging |
1.00 |
R2178:Pappa
|
UTSW |
4 |
65269924 |
missense |
probably benign |
0.00 |
R2504:Pappa
|
UTSW |
4 |
65099126 |
nonsense |
probably null |
|
R4043:Pappa
|
UTSW |
4 |
65232824 |
missense |
probably benign |
0.05 |
R4289:Pappa
|
UTSW |
4 |
65074100 |
missense |
probably benign |
0.19 |
R4415:Pappa
|
UTSW |
4 |
65223532 |
missense |
probably benign |
0.00 |
R4529:Pappa
|
UTSW |
4 |
65149419 |
missense |
probably benign |
|
R4620:Pappa
|
UTSW |
4 |
65245265 |
missense |
probably benign |
0.43 |
R4657:Pappa
|
UTSW |
4 |
65233033 |
splice site |
probably null |
|
R4658:Pappa
|
UTSW |
4 |
65233033 |
splice site |
probably null |
|
R5074:Pappa
|
UTSW |
4 |
65123365 |
missense |
probably benign |
0.15 |
R5200:Pappa
|
UTSW |
4 |
65074076 |
missense |
probably damaging |
1.00 |
R5420:Pappa
|
UTSW |
4 |
65254017 |
critical splice donor site |
probably null |
|
R5469:Pappa
|
UTSW |
4 |
65123389 |
missense |
probably benign |
0.01 |
R5651:Pappa
|
UTSW |
4 |
65074589 |
missense |
probably damaging |
0.99 |
R5725:Pappa
|
UTSW |
4 |
65107647 |
missense |
probably damaging |
1.00 |
R5941:Pappa
|
UTSW |
4 |
65232830 |
missense |
possibly damaging |
0.52 |
R6002:Pappa
|
UTSW |
4 |
65215645 |
missense |
probably damaging |
0.99 |
R6252:Pappa
|
UTSW |
4 |
65107649 |
missense |
probably benign |
0.02 |
R6303:Pappa
|
UTSW |
4 |
65122891 |
missense |
probably damaging |
1.00 |
R6322:Pappa
|
UTSW |
4 |
65232896 |
missense |
probably damaging |
1.00 |
R6431:Pappa
|
UTSW |
4 |
65074701 |
missense |
probably damaging |
1.00 |
R6462:Pappa
|
UTSW |
4 |
65043128 |
missense |
probably damaging |
0.98 |
R6484:Pappa
|
UTSW |
4 |
65232896 |
missense |
probably damaging |
1.00 |
R6537:Pappa
|
UTSW |
4 |
65215519 |
missense |
probably damaging |
0.99 |
R6578:Pappa
|
UTSW |
4 |
65074374 |
missense |
possibly damaging |
0.48 |
R6704:Pappa
|
UTSW |
4 |
65123161 |
missense |
probably damaging |
1.00 |
R6789:Pappa
|
UTSW |
4 |
65099278 |
missense |
probably damaging |
1.00 |
R7023:Pappa
|
UTSW |
4 |
65269955 |
missense |
probably benign |
0.00 |
R7139:Pappa
|
UTSW |
4 |
65107687 |
missense |
probably benign |
0.30 |
R7158:Pappa
|
UTSW |
4 |
65123104 |
missense |
possibly damaging |
0.94 |
R7165:Pappa
|
UTSW |
4 |
65180110 |
missense |
probably damaging |
1.00 |
R7196:Pappa
|
UTSW |
4 |
65242128 |
splice site |
probably null |
|
R7410:Pappa
|
UTSW |
4 |
65253956 |
missense |
probably damaging |
1.00 |
R7457:Pappa
|
UTSW |
4 |
65107503 |
missense |
probably damaging |
1.00 |
R7506:Pappa
|
UTSW |
4 |
65149419 |
missense |
probably benign |
0.00 |
R7546:Pappa
|
UTSW |
4 |
65074352 |
missense |
possibly damaging |
0.48 |
R7975:Pappa
|
UTSW |
4 |
65212705 |
missense |
probably damaging |
0.96 |
R8111:Pappa
|
UTSW |
4 |
65180229 |
missense |
probably damaging |
0.99 |
R8260:Pappa
|
UTSW |
4 |
65234419 |
missense |
probably damaging |
0.99 |
R8347:Pappa
|
UTSW |
4 |
65245302 |
missense |
probably damaging |
1.00 |
R8520:Pappa
|
UTSW |
4 |
65254001 |
missense |
probably benign |
0.01 |
R8812:Pappa
|
UTSW |
4 |
65123166 |
missense |
possibly damaging |
0.94 |
R8815:Pappa
|
UTSW |
4 |
65099347 |
missense |
probably benign |
0.00 |
R9008:Pappa
|
UTSW |
4 |
65074426 |
missense |
probably damaging |
1.00 |
R9162:Pappa
|
UTSW |
4 |
65123040 |
missense |
probably damaging |
1.00 |
R9170:Pappa
|
UTSW |
4 |
65258962 |
missense |
probably damaging |
1.00 |
R9205:Pappa
|
UTSW |
4 |
65074612 |
missense |
possibly damaging |
0.66 |
R9336:Pappa
|
UTSW |
4 |
65042918 |
missense |
unknown |
|
R9389:Pappa
|
UTSW |
4 |
65099125 |
missense |
probably damaging |
1.00 |
R9781:Pappa
|
UTSW |
4 |
65043104 |
missense |
possibly damaging |
0.89 |
RF006:Pappa
|
UTSW |
4 |
65242110 |
missense |
probably benign |
0.00 |
RF020:Pappa
|
UTSW |
4 |
65123282 |
missense |
possibly damaging |
0.77 |
X0058:Pappa
|
UTSW |
4 |
65074469 |
missense |
probably damaging |
1.00 |
X0060:Pappa
|
UTSW |
4 |
65043178 |
missense |
probably benign |
0.00 |
Z1177:Pappa
|
UTSW |
4 |
65225995 |
missense |
probably damaging |
1.00 |
|
Mode of Inheritance |
Unknown |
Local Stock | |
Repository | |
Last Updated |
2019-09-04 9:25 PM
by Anne Murray
|
Record Created |
2019-05-18 2:02 PM
by Bruce Beutler
|
Record Posted |
2019-05-22 |
Phenotypic Description |
The untersuchen phenotype was identified among G3 mice of the pedigree R2127, some of which showed reduced body weights compared to wild-type littermates (Figure 1).
|
Nature of Mutation |
Whole exome HiSeq sequencing of the G1 grandsire identified 94 mutations. The body weight phenotype was linked to mutations in several genes. However, the mutation in Pappa was presumed causative as the body weight phenotype in the untersuchen mice mimics that of mice expressing other mutant Pappa alleles (see MGI). The mutation in Pappa is a T to A transversion at base pair 65,297,257 (v38) on chromosome 4, or base pair 173,084 in the GenBank genomic region NC_000070. Linkage was found with a recessive model of inheritance, wherein one variant homozygote departed phenotypically from 11 homozygous reference mice and 12 heterozygous mice with a P value of 0.000366 (Figure 2). The mutation corresponds to residue 3,768 in the mRNA sequence NM_021362 within exon 13 of 22 total exons.
3753 CTAGGCCTCCATGTCTTGAGCTGCAGAAACAAT
1129 -L--G--L--H--V--L--S--C--R--N--N-
|
The mutated nucleotide is indicated in red. The mutation results in a leucine to methionine substitution at position 1,134 (L1134M) in the PAPP-A protein, and is strongly predicted by Polyphen-2 to be damaging (score = 1.000).
|
Illustration of Mutations in
Gene & Protein |
|
---|
Protein Prediction |
Pappa encodes pregnancy-associated plasma protein A (PAPP-A; alternatively, IGFBP4 protease or differentially expressed in placenta 1 [DIPLA1]), a member of the pappalysin subfamily of the metzincin protease family along with PAPP-A2 (see the record for Lilliputian) and ulilysin. PAPP-A is initially translated as a 1,624 proprotein; cleavage of the signal peptide and the propeptide (amino acids 23-80) generates the mature 1,544 amino acid peptide (1). PAPP-A has several domains, including a signaling peptide (amino acids 1-22), a laminin G-like domain, three Lin12/Notch repeats (LNRs), a metalloprotease region (amino acids 272-583), and five complement control protein (CCP) domains (alternatively, short consensus repeat [SCR] or Sushi domains; amino acids 1210-1279, 1280-1341, 1342-1409, 1410-1470, and 1473-1553) (Figure 3) (2). The untersuchen mutation results in a leucine to methionine substitution at position 1,134 (L1134M); Leu1134 is within an undefined region before the first CCP domain. Please see the record caer for more information about Pappa.
|
Putative Mechanism | PAPP-A is a secreted metalloproteinase that cleaves insulin-like growth factor binding protein 2 (IGFBP2), IGFBP4, and IGFBP5. The IGFBPs regulate the IGF-I signaling pathways by binding IGF-I. IGFBP5 also has IGF-I-independent functions. IGFPB5 is able to bind its putative receptor to enter the cytoplasm and subsequently interact with, and regulate, other proteins. IGFBPs are carrier proteins that regulate the bioavailability of insulin-like growth factors (IGFs) by prolonging their-half-life and circulation turnover. IGFs are essential for the regulation of growth and development by influencing the proliferation, differentiation, and apoptosis of osteoblasts (3;4). IGFs bind to two types of receptors, IGF-IR and IGF-IIR, subsequently activating downstream tyrosine kinase pathways. In IGF-I-associated signaling, both the IRS-1/phosphoinositide 3-kinase/serine–threonine kinase pathway and the Ras/mitogen-activated protein kinase/extracellular signal-regulated kinase pathway are activated, which subsequently promote cell proliferation, tissue differentiation, and protection from apoptosis. Pappa-deficient (Pappa-/-) mice showed reduced body sizes and weights (60 to 70% of wild-type mice) as well as delayed bone ossification (5). The phenotype of the untersuchen mice mimics that of the Pappa-/- mice, indicating loss of PAPP-A function. PAPP-A functions as a growth-promoting enzyme through its role in cleaving IGFBPs (and subsequent release of bioactive IGF). IGFBP4 cleavage is required to activate most, if not all, IGF2-mediated growth-promoting activity.
|
Primers |
PCR Primer
untersuchen_pcr_F: AAGTCCCTGTAGCAACGTGG
untersuchen_pcr_R: GGTTGCTTACCTCTGCTCAG
Sequencing Primer
untersuchen_seq_F: CCATAGATGAGGGTGGCCATTTG
untersuchen_seq_R: AGGGCTGTAGGTCTCTCCTC
|
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 544 nucleotides is amplified (chromosome 4, + strand):
1 aagtccctgt agcaacgtgg gagggaggga gggagagcca tagatgaggg tggccatttg 61 ggggagggag agactgtgag cattaggtga aggaaagctt aggggccagg gaggcagggg 121 gattggaaag atttggaagc acggaaaaaa ggcagacaaa gaaactttca atcaaactca 181 aaatcctcac tggccgccca atggtgcttt tgtggagtga catgaaaggg tcttggggct 241 tcgggaagag cccgttgggg agggcgtgac tggccttgag taatgagcct ggcccctcgg 301 ccctctgtct cctctgacag gcctccatgt cttgagctgc agaaacaatc ccctgattat 361 ccctgtggtc catgacctca gccagccctt ctaccacagc caggcggtac atgtgagctt 421 cagttcgccc ctggtcgcca tctcgggggt ggccctccgc tctttcgaca actttgaccc 481 cgtcaccctg agcagctgcc agagaggaga gacctacagc cctgctgagc agaggtaagc 541 aacc
Primer binding sites are underlined and the sequencing primers are highlighted; the mutated nucleotide is shown in red. |
References | 1. Haaning, J., Oxvig, C., Overgaard, M. T., Ebbesen, P., Kristensen, T., and Sottrup-Jensen, L. (1996) Complete cDNA Sequence of the Preproform of Human Pregnancy-Associated Plasma Protein-A. Evidence for Expression in the Brain and Induction by cAMP. Eur J Biochem. 237, 159-163.
2. Kristensen, T., Oxvig, C., Sand, O., Moller, N. P., and Sottrup-Jensen, L. (1994) Amino Acid Sequence of Human Pregnancy-Associated Plasma Protein-A Derived from Cloned cDNA. Biochemistry. 33, 1592-1598.
4. Mohan, S., Richman, C., Guo, R., Amaar, Y., Donahue, L. R., Wergedal, J., and Baylink, D. J. (2003) Insulin-Like Growth Factor Regulates Peak Bone Mineral Density in Mice by both Growth Hormone-Dependent and -Independent Mechanisms. Endocrinology. 144, 929-936.
5. Conover, C. A., Bale, L. K., Overgaard, M. T., Johnstone, E. W., Laursen, U. H., Fuchtbauer, E. M., Oxvig, C., and van Deursen, J. (2004) Metalloproteinase Pregnancy-Associated Plasma Protein A is a Critical Growth Regulatory Factor during Fetal Development. Development. 131, 1187-1194.
|
Science Writers | Anne Murray |
Illustrators | Diantha La Vine |
Authors | Zhao Zhang and Bruce Beutler |