Summary
The Lesch-Nyhan syndrome is a severe X chromosome-linked human disease caused by a virtual absence of hypoxanthine-guanine phosphoribosyltransferase (HPRT) activity. A partial deficiency in the activity of this enzyme can result in gouty arthritis. To determine the genetic basis for reduction or loss of enzyme activity, we have amplified and sequenced the coding region of HPRT cDNA from four patients: one with LeschNyhan syndrome (HPRTPerth) and three with partial deficiencies of HPRT activity, which have been designated HPRTUrangan, HPRTSwan and HPRTToowong. In all four patients, the only mutation identified was a single base substitution in exons 2 or 3 of the coding region, which in each case predicts a single amino acid substitution in the translated protein. Each base change was confirmed by allele-specific amplification of the patient's genomic DNA. It is interesting to note that the mutation found for HPRTPerth is identical to that reported for HPRTFlint. It appears that the two mutations are de novo events.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Argos P, Hanei M, Wilson JM, Kelley WN (1983) A possible nucleotide-binding domain in the tertiary fold of phosphoribosyltransferases. J Biol Chem 258:6450–6457
Chirgwin JM, Przybyla AE, MacDonald RJ, Rutter WJ (1979) Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18:5294–5299
Craig SP, McKerrow JH, Newport GR, Wang CC (1988) Analysis of cDNA encoding the hypoxanthine-guanine phosphoribosyltransferase (HGPRTase) of Schistosoma mansoni; a putative target for chemotherapy. Nucleic Acids Res 16/14:7087–7107
Davidson B, Pashmforoush M, Kelley WN, Palella TD (1988a) Genetic basis of hypoxanthine guanine phosphoribosyltransferase deficiency in a patient with the Lesch-Nyhan syndrome (HPRTFlint). Gene 63:331–336
Davidson B, Chin SJ, Wilson J, Kelley W, Palella T (1988b) Hypoxanthine-guanine phosphoribosyltransferase — genetic evidence for identical mutations in two partially deficient subjects. J Clin Invest 82:2164–2167
Davidson BL, Tarle SA, Palella TD, Kelley WN (1989) Molecular basis of hypoxanthine-guanine phosphoribosyltransferase deficiency in ten subjects determine by direct sequencing of amplified transcripts. J Clin Invest 84:342–346
Davis LG, Dibner MD, Battey JF (1986) Basis methods in molecular biology. Elsevier Science, New York
Gibbs RA, Nguyen PH, McBride LJ, Koepf SM, Caskey CT (1989) Identification of mutations leading to the Lesch-Nyhan syndrome by automated direct DNA sequencing of in vitro amplified cDNA. Proc Natl Acad Sci USA 86:1919–1923
Gibbs RA, Nguyen P-N, Edwards A, Civitello AB, Caskey CT (1990) Multiplex DNA deletion and exon sequencing of the hypoxanthine phosphoribosyltransferase gene in Lesch-Nyhan families. Genomics 7:235–244
Gordon RB, Emmerson BT, Stout JT, Caskey CT (1987) Molecular studies of hypoxanthine-guanine phosphoribosyltransferase mutations in six Australian families. Aust NZ J Med 17:424–429
Gordon RB, Sculley DG, Dawson PA, Beacham IF, Emmerson BT (1990) Identification of a single nucleotide substitution of in vitro amplified cDNA from a patient with partial HPRT-deficiency (HPRTBrisbane). J Inherited Metab Dis 13:692–700
Jolly DJ, Okayama H, Berg P, Esty AC, Filpula D, Bohlen P, Johnson GG, Shively JE, Hunkapillar T, Friedmann T (1983) Isolation and characterisation of a full-length expressible cDNA for human hypoxanthine phosphoribosyltransferase. Proc Natl Acad Sci USA 80:477–481
Keough DT, Gordon RB, Jersey J de, Emmerson BT (1988) Biochemical basis of HPRT deficiency in nine families. J Inherited Metab Dis 11:229–238
Kogan SC, Doherty M, Gitschier J (1987) An improved method for prenatal diagnosis of genetic diseases by analysis of amplified DNA sequences. N Engl J Med 317:985–990
Lesch M, Nyhan WL (1964) A familial disorder of uric acid metabolism and central nervous system function. Am J Med 36:561–570
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
Saiki RK, Gelfand DH, Stoffel SS, Scharf SJ, Higuchi R, Horn T, Mullis KB, Erlich HE (1988) Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239:487–491
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Sculley, D.G., Dawson, P.A., Beacham, I.R. et al. Hypoxanthine-guanine phosphoribosyltransferase deficiency: analysis of HPRT mutations by direct sequencing and allele-specific amplification. Hum Genet 87, 688–692 (1991). https://doi.org/10.1007/BF00201727
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00201727