Description:
PHENYLKETONURIA
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Repository
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NIGMS Human Genetic Cell Repository
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| Subcollection |
Heritable Diseases |
| Class |
Disorders of Amino Acid Metabolism |
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Cell Type
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Fibroblast
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Transformant
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Untransformed
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Race
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White
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Family Member
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1
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Relation to Proband
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proband
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Confirmation
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Clinical summary/Case history
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Species
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Homo sapiens
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Common Name
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Human
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Remarks
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| PDL at Freeze |
4.95 |
| Passage Frozen |
4 |
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| IDENTIFICATION OF SPECIES OF ORIGIN |
Species of Origin Confirmed by Nucleoside Phosphorylase Isoenzyme Electrophoresis |
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| Gene |
PAH |
| Chromosomal Location |
12q24.1 |
| Allelic Variant 1 |
261600.0039; PHENYLKETONURIA |
| Identified Mutation |
PHE299CYS; The mutant haplotype 8 occurs relatively frequently in Norwegian PKU patients (comprising 6% of mutant genes), whereas it is rare among other European PKU patients. Normal haplotype 8 genes have not been observed in any European population. Eiken et al. (1992) found that all mutant haplotype 8 chromosomes carried the phe299-to-cys mutation described briefly by Okano et al. (1989). A patient homozygous for the F299C mutation manifested severe PKU.
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| Gene |
PAH |
| Chromosomal Location |
12q24.1 |
| Allelic Variant 2 |
261600.0002; PHENYLKETONURIA |
| Identified Mutation |
ARG408TRP; DiLella et al. (1987) reported the molecular lesion associated with the RFLP haplotype-2 mutant allele. This defect is caused by a CGG-to-TGG transition in exon 12, resulting in an amino acid substitution (arg-to-trp) at residue 408 of PAH. Direct hybridization analysis of the point mutation using a specific oligonucleotide probe demonstrated that this mutation is in linkage disequilibrium with RFLP haplotype-2 alleles that make up about 20% of mutant PAH genes. This is presumably another example of CpG mutation. Woo (1988) provided a collation of the 43 RFLP haplotypes at the PAH locus identified to date. Ninety percent of all mutant alleles in Danes are associated with only 4 haplotypes, of which 2 had been fully characterized at the molecular level. The haplotypes are based on the combined pattern of presence or absence of sites of cutting by 7 restriction enzymes (BglIII, PvuII, EcoRI, MspI, XmnI, HindIII, and EcoRV), of which one, PvuII, has 2 cut sites. The GT-to-AT transition at the canonical splice donor site of intron 12, causing skipping of the preceding exon during RNA splicing, is associated with a mutant haplotype 3. The missense mutation involving an arginine-to-tryptophan substitution at residue number 408 of the enzyme is associated with mutant haplotype 2. Both mutant alleles are in linkage disequilibrium with the corresponding RFLP haplotypes throughout Europe, suggesting that 2 mutational events occurred on background chromosomes of the 2 haplotypes, followed by spread and expansion in the Caucasian population. In French Canadians, John et al. (1990) found that the arg408-to-trp mutation in exon 12 is associated with haplotype 1; in other populations, it occurs on haplotype 2. A CpG dinucleotide is involved in this mutation, compatible with a recurrent mutation, although gene conversion or a single recombination between haplotypes 2 and 1 is possible. Kalaydjieva et al. (1991) found this mutation in high frequency in Bulgaria, Lithuania, and eastern Germany, where it occurred on haplotype 2. Pooling of data on European populations suggested a Balto-Slavic origin of the arg408-to-trp defect, with an east-west cline in its frequency. Tsai et al. (1990) found this mutation in Chinese on a different haplotype, namely, no. 44. John et al. (1990) presented a tabulation of 20 PAH mutations showing 3 instances of putative recurrent mutation. Jaruzelska et al. (1991) found that haplotype 2 was most frequently (62%) associated with PKU alleles in Poland, where in the western part of the country the frequency of PKU is 1 in 5,000 live births. Furthermore, the arg408-to-trp mutation was in complete linkage disequilibrium with this haplotype. Similar observations have been made in other eastern European countries such as the former German Democratic Republic, Czechoslovakia, and Hungary. Zygulska et al. (1991) found similar results in southern Poland. Zygulska et al. (1991) found the arg408-to-trp mutation in 25 of 44 chromosomes from 22 unrelated Polish families with at least 1 PKU child. In 24 of these, mutation was on haplotype 2. A different mutation in the same codon, arg408-to-gln (261600.0038), has been described. Recurrent mutations in the 408 codon appear to occur; at least 2 different mutations (at least mutations on different RFLP haplotype background) have been identified in Chinese (Lin et al., 1992). Codon 408 (CGG) contains a CpG hotspot (Ramus et al., 1992). The R408W mutation is a CGG-to-TGG change in the coding strand; the R408Q mutation (261600.0038) is a GCC-to-GTC change in the noncoding strand. Ivaschenko and Baranov (1993) described a rapid and efficient PCR/StyI test for identification of this mutation.
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| Remarks |
Clinically affected; pyloric stenosis; normal level of dihydropteridine reductase in skin fibroblasts; donor subject is a compound heterozygote: one allele has a T>G transversion at nucleotide 896 in exon 8 of the PAH gene [896T>G] resulting in a substitution of cysteine for phenylalanine at codon 299 [Phe299Cys (F299C)] and a second allele has a C>T transition at nucleotide 1222 in exon 11 of the PAH gene [1222C>T] resulting in a substitution of tryptophan for arginine at codon 408 [Arg408Trp (R408W)]. Same subject as GM27578 (stem cell). |
| Veleva D, Ay M, Ovchinnikov DA, Prowse ABJ, Menezes MJ, Nafisinia M, Generation of fibroblast-derived induced pluripotent stem cell (iPSC) lines from two paediatric patients with phenylketonuria Stem cell research77:103405 2024 |
| PubMed ID: 38555716 |
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| Veleva D, Ay M, Ovchinnikov DA, Prowse ABJ, Menezes MJ, Nafisinia M, Generation of two lymphoblastoid-derived induced pluripotent stem cell (iPSC) lines from patients with phenylketonuria Stem cell research77:103407 2024 |
| PubMed ID: 38552357 |
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| Yeeok Kang, Seong-Hyeuk Nam, Kyung Sun Park, Yoonjung Kim, Jong-Won Kim, Eunjung Lee, Jung Min Ko, Kyung-A Lee and Inho ParkEmail, DeviCNV: detection and visualization of exon-level copy number variants in targeted next-generation sequencing data BMC Bioinformatics19:103407 2018 |
| PubMed ID: 30326846 |
| Passage Frozen |
4 |
| Split Ratio |
1:4 |
| Temperature |
37 C |
| Percent CO2 |
5% |
| Percent O2 |
AMBIENT |
| Medium |
Eagle's Minimum Essential Medium with Earle's salts and non-essential amino acids with 2mM L-glutamine or equivalent |
| Serum |
15% fetal bovine serum Not inactivated |
| Supplement |
- |
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