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. 1990 Jan;46(1):18–25.

Missense mutations associated with RFLP haplotypes 1 and 4 of the human phenylalanine hydroxylase gene.

Y Okano 1, T Wang 1, R C Eisensmith 1, B Steinmann 1, R Gitzelmann 1, S L Woo 1
PMCID: PMC1683547  PMID: 1967207

Abstract

We report missense mutations associated with haplotype 1 and haplotype 4 alleles of the human phenylalanine hydroxylase (PAH) gene. Individual exon-containing regions were amplified by polymerase chain reaction from genomic DNA of a PKU patient who was a haplotype 1/4 compound heterozygote. The amplified DNA fragments were subcloned into M13 for sequence analysis. Missense mutations were observed in exons 5 and 7, resulting in the substitution of Arg by Gln at residues 158 and 261 of the enzyme, respectively. Expression analysis in heterozygous mammalian cells after site-directed mutagenesis demonstrated that the Arg158-to-Gln158 mutation is a PKU mutation, whereas the Arg261-to-Gln261 mutation is apparently silent in the assay system. Hybridization analysis using allele-specific oligonucleotide probes demonstrated that the Arg158-to-Gln158 mutation is present in two of six mutant haplotype 4 alleles among the Swiss and constitutes about 40% of all mutant haplotype 4 alleles in the European population. The mutation is not present in normal alleles or in any mutant alleles of other haplotypes. The results provide conclusive evidence that there is linkage disequilibrium between mutation and haplotype in the PAH gene and that multiple mutations have occurred in the PAH gene of a prevalent haplotype among Caucasians.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aulehla-Scholz C., Vorgerd M., Sautter E., Leupold D., Mahlmann R., Ullrich K., Olek K., Horst J. Phenylketonuria: distribution of DNA diagnostic patterns in German families. Hum Genet. 1988 Apr;78(4):353–355. doi: 10.1007/BF00291734. [DOI] [PubMed] [Google Scholar]
  2. Chakraborty R., Lidsky A. S., Daiger S. P., Güttler F., Sullivan S., Dilella A. G., Woo S. L. Polymorphic DNA haplotypes at the human phenylalanine hydroxylase locus and their relationship with phenylketonuria. Hum Genet. 1987 May;76(1):40–46. doi: 10.1007/BF00283048. [DOI] [PubMed] [Google Scholar]
  3. Daiger S. P., Chakraborty R., Reed L., Fekete G., Schuler D., Berenssi G., Nasz I., Brdicka R., Kamarýt J., Pijácková A. Polymorphic DNA haplotypes at the phenylalanine hydroxylase (PAH) locus in European families with phenylketonuria (PKU). Am J Hum Genet. 1989 Aug;45(2):310–318. [PMC free article] [PubMed] [Google Scholar]
  4. DiLella A. G., Huang W. M., Woo S. L. Screening for phenylketonuria mutations by DNA amplification with the polymerase chain reaction. Lancet. 1988 Mar 5;1(8584):497–499. doi: 10.1016/s0140-6736(88)91295-0. [DOI] [PubMed] [Google Scholar]
  5. DiLella A. G., Kwok S. C., Ledley F. D., Marvit J., Woo S. L. Molecular structure and polymorphic map of the human phenylalanine hydroxylase gene. Biochemistry. 1986 Feb 25;25(4):743–749. doi: 10.1021/bi00352a001. [DOI] [PubMed] [Google Scholar]
  6. DiLella A. G., Marvit J., Brayton K., Woo S. L. An amino-acid substitution involved in phenylketonuria is in linkage disequilibrium with DNA haplotype 2. 1987 May 28-Jun 3Nature. 327(6120):333–336. doi: 10.1038/327333a0. [DOI] [PubMed] [Google Scholar]
  7. DiLella A. G., Marvit J., Lidsky A. S., Güttler F., Woo S. L. Tight linkage between a splicing mutation and a specific DNA haplotype in phenylketonuria. 1986 Aug 28-Sep 3Nature. 322(6082):799–803. doi: 10.1038/322799a0. [DOI] [PubMed] [Google Scholar]
  8. DiLella A. G., Woo S. L. Hybridization of genomic DNA to oligonucleotide probes in the presence of tetramethylammonium chloride. Methods Enzymol. 1987;152:447–451. doi: 10.1016/0076-6879(87)52052-3. [DOI] [PubMed] [Google Scholar]
  9. GUTHRIE R., SUSI A. A SIMPLE PHENYLALANINE METHOD FOR DETECTING PHENYLKETONURIA IN LARGE POPULATIONS OF NEWBORN INFANTS. Pediatrics. 1963 Sep;32:338–343. [PubMed] [Google Scholar]
  10. Güttler F. Hyperphenylalaninemia: diagnosis and classification of the various types of phenylalanine hydroxylase deficiency in childhood. Acta Paediatr Scand Suppl. 1980;280:1–80. [PubMed] [Google Scholar]
  11. Güttler F., Ledley F. D., Lidsky A. S., DiLella A. G., Sullivan S. E., Woo S. L. Correlation between polymorphic DNA haplotypes at phenylalanine hydroxylase locus and clinical phenotypes of phenylketonuria. J Pediatr. 1987 Jan;110(1):68–71. doi: 10.1016/s0022-3476(87)80290-1. [DOI] [PubMed] [Google Scholar]
  12. Kwok S. C., Ledley F. D., DiLella A. G., Robson K. J., Woo S. L. Nucleotide sequence of a full-length complementary DNA clone and amino acid sequence of human phenylalanine hydroxylase. Biochemistry. 1985 Jan 29;24(3):556–561. doi: 10.1021/bi00324a002. [DOI] [PubMed] [Google Scholar]
  13. Ledley F. D., Grenett H. E., DiLella A. G., Kwok S. C., Woo S. L. Gene transfer and expression of human phenylalanine hydroxylase. Science. 1985 Apr 5;228(4695):77–79. doi: 10.1126/science.3856322. [DOI] [PubMed] [Google Scholar]
  14. Ledley F. D., Koch R., Jew K., Beaudet A., O'Brien W. E., Bartos D. P., Woo S. L. Phenylalanine hydroxylase expression in liver of a fetus with phenylketonuria. J Pediatr. 1988 Sep;113(3):463–468. doi: 10.1016/s0022-3476(88)80629-2. [DOI] [PubMed] [Google Scholar]
  15. Lichter-Konecki U., Konecki D. S., DiLella A. G., Brayton K., Marvit J., Hahn T. M., Trefz F. K., Woo S. L. Phenylalanine hydroxylase deficiency caused by a single base substitution in an exon of the human phenylalanine hydroxylase gene. Biochemistry. 1988 Apr 19;27(8):2881–2885. doi: 10.1021/bi00408a032. [DOI] [PubMed] [Google Scholar]
  16. Lichter-Konecki U., Schlotter M., Konecki D. S., Labeit S., Woo S. L., Trefz F. K. Linkage disequilibrium between mutation and RFLP haplotype at the phenylalanine hydroxylase locus in the German population. Hum Genet. 1988 Apr;78(4):347–352. doi: 10.1007/BF00291733. [DOI] [PubMed] [Google Scholar]
  17. Lidsky A. S., Güttler F., Woo S. L. Prenatal diagnosis of classic phenylketonuria by DNA analysis. Lancet. 1985 Mar 9;1(8428):549–551. doi: 10.1016/s0140-6736(85)91208-5. [DOI] [PubMed] [Google Scholar]
  18. Lidsky A. S., Ledley F. D., DiLella A. G., Kwok S. C., Daiger S. P., Robson K. J., Woo S. L. Extensive restriction site polymorphism at the human phenylalanine hydroxylase locus and application in prenatal diagnosis of phenylketonuria. Am J Hum Genet. 1985 Jul;37(4):619–634. [PMC free article] [PubMed] [Google Scholar]
  19. Lyonnet S., Caillaud C., Rey F., Berthelon M., Frézal J., Rey J., Munnich A. Molecular genetics of phenylketonuria in Mediterranean countries: a mutation associated with partial phenylalanine hydroxylase deficiency. Am J Hum Genet. 1989 Apr;44(4):511–517. [PMC free article] [PubMed] [Google Scholar]
  20. Marvit J., DiLella A. G., Brayton K., Ledley F. D., Robson K. J., Woo S. L. GT to AT transition at a splice donor site causes skipping of the preceding exon in phenylketonuria. Nucleic Acids Res. 1987 Jul 24;15(14):5613–5628. doi: 10.1093/nar/15.14.5613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  22. Rey F., Berthelon M., Caillaud C., Lyonnet S., Abadie V., Blandin-Savoja F., Feingold J., Saudubray J. M., Frézal J., Munnich A. Clinical and molecular heterogeneity of phenylalanine hydroxylase deficiencies in France. Am J Hum Genet. 1988 Dec;43(6):914–921. [PMC free article] [PubMed] [Google Scholar]
  23. Riess O., Michel A., Speer A., Meiske W., Cobet G., Coutelle C. Linkage disequilibrium between RFLP haplotype 2 and the affected PAH allele in PKU families from the Berlin area of the German Democratic Republic. Hum Genet. 1988 Apr;78(4):343–346. doi: 10.1007/BF00291732. [DOI] [PubMed] [Google Scholar]
  24. Robson K. J., Chandra T., MacGillivray R. T., Woo S. L. Polysome immunoprecipitation of phenylalanine hydroxylase mRNA from rat liver and cloning of its cDNA. Proc Natl Acad Sci U S A. 1982 Aug;79(15):4701–4705. doi: 10.1073/pnas.79.15.4701. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Scriver C. R., Kaufman S., Woo S. L. Mendelian hyperphenylalaninemia. Annu Rev Genet. 1988;22:301–321. doi: 10.1146/annurev.ge.22.120188.001505. [DOI] [PubMed] [Google Scholar]
  26. Silva A. J., White R. Inheritance of allelic blueprints for methylation patterns. Cell. 1988 Jul 15;54(2):145–152. doi: 10.1016/0092-8674(88)90546-6. [DOI] [PubMed] [Google Scholar]
  27. Sullivan S. E., Moore S. D., Connor J. M., King M., Cockburn F., Steinmann B., Gitzelmann R., Daiger S. P., Woo S. L. Haplotype distribution of the human phenylalanine hydroxylase locus in Scotland and Switzerland. Am J Hum Genet. 1989 May;44(5):652–659. [PMC free article] [PubMed] [Google Scholar]
  28. Wong G. G., Witek J. S., Temple P. A., Wilkens K. M., Leary A. C., Luxenberg D. P., Jones S. S., Brown E. L., Kay R. M., Orr E. C. Human GM-CSF: molecular cloning of the complementary DNA and purification of the natural and recombinant proteins. Science. 1985 May 17;228(4701):810–815. doi: 10.1126/science.3923623. [DOI] [PubMed] [Google Scholar]
  29. Woo S. L., Lidsky A. S., Güttler F., Chandra T., Robson K. J. Cloned human phenylalanine hydroxylase gene allows prenatal diagnosis and carrier detection of classical phenylketonuria. Nature. 1983 Nov 10;306(5939):151–155. doi: 10.1038/306151a0. [DOI] [PubMed] [Google Scholar]
  30. Woo S. L. Molecular basis and population genetics of phenylketonuria. Biochemistry. 1989 Jan 10;28(1):1–7. doi: 10.1021/bi00427a001. [DOI] [PubMed] [Google Scholar]
  31. Zoller M. J., Smith M. Oligonucleotide-directed mutagenesis of DNA fragments cloned into M13 vectors. Methods Enzymol. 1983;100:468–500. doi: 10.1016/0076-6879(83)00074-9. [DOI] [PubMed] [Google Scholar]

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