Human Genetics

, Volume 79, Issue 1, pp 39–43 | Cite as

Identification of a single nucleotide change in a mutant gene for hypoxanthine-guanine phosphoribosyltransferase (HPRTAnn Arbor)

  • Shin Fujimori
  • Yuji Hidaka
  • Beverly L. Davidson
  • Thomas D. Palella
  • William N. Kelley
Original Investigations


HPRTAnn Arbor is a variant of hypoxanthine (guanine) phosphoribosyl-transferase (HPRT: EC, which was identified in two brothers with hyperuricemia and nephrolithiasis. In previous studies, this mutant enzyme was characterized by an increased Km for both substrates, a normal Vmax, a decreased intracellular concentration of enzyme protein, a normal subunit molecular weight and an acidic isoelectric point under native isoelectric focusing conditions. We have cloned a full-length cDNA for HPRTAnn Arbor and determined its complete nucleotide sequence. A single nucleotide change (T→G) at nucleotide position 396 has been identified. This transversion predicts an amino acid substitution from isoleucine (ATT) to methionine (ATG) in codon 132, which is located within the putative 5′-phosphoribosyl-1-pyrophosphate (PRPP)-binding site of HPRT.


Codon Methionine Guanine Amino Acid Substitution Isoelectric Point 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Argos P, Hanei M, Wilson JM, Kelley WN (1983) A possible nucleotide-binding domain in the tertiary fold of phosphoribosyltransferase. J Biol Chem 258: 6450–6457Google Scholar
  2. Aviv H, Leder P (1972) Purification of biologically active globin mRNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci USA 69: 1408–1412Google Scholar
  3. Benton WD, Davis RW (1977) Screening λgt recombinant clones by hybridization to single plaques in situ. Science 196: 180–182Google Scholar
  4. Biggin MD, Gibson TJ, Hong GF (1983) Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci USA 80: 3963–3965Google Scholar
  5. Caskey CT (1987) Disease diagnosis by recombinant DNA methods. Science 236: 1223–1229Google Scholar
  6. Chinault AC, Caskey CT (1984) The hypoxanthine phosphoribosyltransferase gene. A model for the study of mutation in mammalian cells. Prog Nucleic Acid Res Mol Biol 31: 295–313Google Scholar
  7. Dretzen G, Bellarn M, Corsi PS, Chambon P (1981) A reliable method for the recovery of DNA fragments from agarose and acrylamide gels. Anal Biochem 112: 295–298Google Scholar
  8. Feinberg AP, Vogelstein B (1984) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 137: 266–267Google Scholar
  9. Fuscoe JC, Fenwich RR, Ledbetter DH, Caskey CT (1983) Deletion and amplification of the HPRT locus in Chinese hamster ovary cells. Mol Cell Biol 3: 1086–1096Google Scholar
  10. Ghangas GS, Milman G (1977) Hypoxanthine phosphoribosyltransferase: two dimensional gels from normal and Lesch-Nyhan hemolysates. Science 196: 1119–1120Google Scholar
  11. Giacomello A, Salermo C (1978) Human hypoxanthine-guanine phosphoribosyl-transferase. Steady state kinetics of the forward and reverse reactions. J Biol Chem 253: 6038–6044Google Scholar
  12. Gibbs RA, Caskey CT (1987) Identification and localization of mutations at the Lesch-Nyhan locus by ribonuclease A cleavage. Science 236:303–305Google Scholar
  13. Gubler U, Hoffman BJ (1983) A simple and very efficient method for generating cDNA libraries. Gene 25:263–269Google Scholar
  14. Hershey HV, Taylor MW (1986) Nucleotide sequence and deduced amino acid sequence of Escherichia coli adenine phosphoribosyltransferase and comparison with other analogous enzymes. Gene 43:287–293Google Scholar
  15. Johnson GG, Eisenberg LR, Migeon BR (1979) Human and mouse hypoxanthine-guanine phosphoribosyltransferase: dimers and tetramers. Science 203:174–176Google Scholar
  16. Johnson GG, Ramage AL, Littlefield JW, Kazazian HH (1982) Hypoxanthine-guanine phosphoribosyltransferase in human erythroid cells. Post-translational modification. Biochemistry 21: 960–966Google Scholar
  17. Jolly DJ, Okayama H, Berg P, Esty AC, Filpula D, Bohlen P, Johnson GG, Shively JE, Hunkapillar T, Friedmann T (1983) Isolation and characterization of a full-length expressible cDNA for human hypoxanthine phosphoribosyltransferase. Proc Natl Acad Sci USA 80:477–481Google Scholar
  18. Kelley WN, Rosenbloom FM, Henderson JF, Seegmiller JE (1967) A specific enzyme defect in gout associated with overproduction of uric acid. Proc Natl Acad Sci USA 57:1735–1739Google Scholar
  19. Maniatis T, Fritsch EF, Sambrook J (eds) (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NYGoogle Scholar
  20. Myers RM, Lavin Z, Maniatis T (1985) Detection of single base substitutions by ribonuclease cleavage at mismatches. Science 230: 1242–1246Google Scholar
  21. Patel PI, Framson PE, Caskey CT, Chinault AC (1986) Fine structure of the human hypoxanthine phosphoribosyltransferase gene. Mol Cell Biol 6:393–403Google Scholar
  22. Polites HG, Marotti KR (1986) A step-wise protocol for cDNA synthesis. Biotechniques 4:514–520Google Scholar
  23. Seegmiller JE, Rosenbloom FM, Kelley WN (1967) Enzyme defect associated with a sex-linked human neurological disorder and excessive purine synthesis. Science 155:1682–1684Google Scholar
  24. Wilson JM, Baugher BW, Mattes PM, Daddona PE, Kelley WN (1982) Human hypoxanthine-guanine phosphoribosyltransferase: demonstration of structural variants in lymphoblastoid cells derived from patients with a deficiency of the enzyme. J Clin Invest 69:706–715Google Scholar
  25. Wilson JM, Young AB, Kelley WN (1983) Hypoxanthine-guanine phosphoribosyltransferase deficiency. N Engl J Med 309:900–910Google Scholar
  26. Wilson JM, Stout JT, Palella TD, Davidson BL, Kelley WN, Caskey CT (1986) A molecular survey of hypoxanthine-guanine phosphoribosyltransferase deficiency in man. J Clin Invest 77:185–195Google Scholar
  27. Winter E, Yamamoto F, Aloguera C, Perucho M (1985) A method to detect and characterize point mutations in transcribed genes: amplification and overexpression of the mutant c-Ki-ras allele in human tumor cells. Proc Natl Acad Sci USA 82:7575–7579Google Scholar
  28. Yang TP, Patel PI, Chinault AC, Stout JT, Jackson LG, Hildebrand BM, Caskey CT (1984) Molecular evidence for new mutation at the HPRT locus in Lesch-Nyhan patients. Nature 310:412–414Google Scholar
  29. Young RA, Davis RW (1983) Efficient isolation of genes by using antibody probes. Proc Natl Acad Sci USA 80:1154–1198Google Scholar
  30. Zannis VI, Gudas LJ, Martin DW (1980) Characterization of the subunit composition of HPRTase from human erythrocytes and cultured fibroblasts. Biochem Genet 18:1–19Google Scholar

Copyright information

© Springer-Verlag 1988

Authors and Affiliations

  • Shin Fujimori
    • 1
  • Yuji Hidaka
    • 1
  • Beverly L. Davidson
    • 2
  • Thomas D. Palella
    • 1
  • William N. Kelley
    • 1
    • 2
  1. 1.Department of Internal MedicineUniversity of MichiganAnn ArborUSA
  2. 2.Department of Biological ChemistryUniversity of MichiganAnn ArborUSA

Personalised recommendations