Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Demonstration of the spf-ash mutation in Spanish patients with ornithine transcarbamylase deficiency of moderate severity

  • 66 Accesses

  • 5 Citations

Abstract

We have found in patients with ornithine transcarbamylase (OTC) deficiency from two Spanish families (A and B), replacement by A of G at the 3′-end of exon 4 of the OTC gene. The same mutation is found in the spf-ash mouse, a rodent model of mild OTC deficiency, causing a neutral R129H mutation and inefficient splicing at the 5′donor site of the exon 4-intron 4 junction, with resultant 4%–7% residual OTC activity. The mutation, detected in our patients using polymerase chain reaction (PCR) amplification of the ten OTC exons, single strand conformation polymorphism (SSCP) analysis and direct sequencing of PCR-amplified exon 4, results in the loss of a unique MspI restriction site which can be used for rapid diagnosis. The mutation was transmitted by the mother in family A and arose de novo in the patient in family B. Residual OTC activity, determined in a male and a female patient, was 1.3% and 3.5% of normal, respectively. Despite this low activity, the surviving patients have developed normally.

This is a preview of subscription content, log in to check access.

References

  1. Briand P, Francois B, Rabier D, Cathelineau L (1982) Ornithine transcarbamylase deficiencies in human males: kinetic and immunochemical classification. Biochim Biophys Acta 704: 100–106

  2. Brusilow SW, Horwich AL (1989) Urea cycle enzymes. In: Scriver CR, Beaudet AL, Sly WS, Valle D (eds) The metabolic basis of inherited disease, 6th edn. McGraw-Hill, New York, pp 629–663

  3. Casanova JL, Pannetier C, Jaulin C, Kourilsky P (1990) Optimal conditions for directly sequencing double-stranded PCR products with Sequenase. Nucleic Acids Res 18: 4028

  4. Dubois N, Cavard C, Chase JF, Kamoun P, Briand P (1988) Compared expression levels of ornithine transcarbamylase and carbamylphosphate synthetase in liver and small intestine of normal and mutant mice. Biochim Biophys Acta 950: 321–328

  5. Garcia-España A, Carbonell J, Rubio V (1989) Carbamoyl phosphate synthetase, ornithine transcarbamylase, and aspartate transcarbamylase activities in the pea ovary. Plant Physiol 90: 1565–1569

  6. Grompe M, Muzny DM, Caskey CT (1989) Scanning detection of mutations in human ornithine transcarbamoylase by chemical mismatch cleavage. Proc Natl Acad Sci USA 86: 5888–5892

  7. Grompe M, Caskey CT, Fenwick RG (1991) Improved molecular diagnostics for ornithine transcarbamylase deficiency. Am J Hum Genet 48: 212–222

  8. Hata A, Tsuzuki T, Shimada K, Takiguchi M, Mori M, Matsuda I (1988) Structure of the human ornithine transcarbamylase gene. J Biochem 103: 302–308

  9. Hodges PE, Rosenberg LE (1989) The spf-ash mouse: a missense mutation in the ornithine transcarbamylase gene also causes aberrant mRNA splicing. Proc Natl Acad Sci USA 86: 4142–4146

  10. Horwich AL, Fenton W, Williams KR, Kalousek F, Kraus JP, Doolittle RF, Koningsberg W, Rosenberg LE (1984) Structure and expression of a complementary DNA for the nuclear coded precursor of human mitocondrial ornithine transcarbamylase. Science 224: 1068–1074

  11. Matsuura T, Hoshide R, Setoyama C, Shimada K, Hase Y, Yanagawa T, Kajita M, Matsuda I (1993) Four novel gene mutations in five Japanese male patients with neonatal or late onset OTC deficiency: application of PCR-single-strand conformation polymorphisms for all exons and adjacent introns. Hum Genet 92: 49–56

  12. Moreno RF, Booth FR, Hung IH, Tilzer LL (1989) Efficient DNA isolation within a single gel barrier tube. Nucleic Acids Res 17: 8393

  13. Nguyen M, Argan C, Lusty CJ, Shore GC (1986) Import and processing of hybrid proteins by mammalian mitochondria in vitro. J Biol Chem 261: 800–805

  14. Nuzum CT, Snodgrass PJ (1976) Multiple assays of the five urea-cycle enzymes in human liver homogenates. In: Grisolía S, Báguena R, Mayor F (eds) The urea cycle. John Wiley & Sons, New York, pp 325–349

  15. Orita M, Iwahana H, Kanazawa H, Hayashi K, Sekiya T (1989) Detection of polymorphisms of human DNA by gel electrophoresis as single-strand conformation polymorphisms. Proc Natl Acad Sci USA 86: 2766–2770

  16. Petty EM, Carstens R, Bale AE (1991) Ornithine transcarbamylase polymorphism detected by PCR introduction of Dra I site. Nucleic Acids Res 19: 690

  17. Shapiro MB, Senapathy P (1987) RNA splice junctions of different classes of eukaryotes: statistics and functional implications in gene expression. Nucleic Acids Res 15: 7155–7174

  18. Sekiya T (1993) Detection of nucleotide sequence changes by single-strand conformation polymorphism analysis. In: Adolph KW (ed) Genome research in molecular medicine and virology. Academic Press, New York, pp 101–113

  19. Strautnieks S, Malcolm S (1993) Novel mutation affecting a splice site in exon 4 of the ornithine carbamoyl transferase gene. Hum Mol Genet 2: 1963–1964

  20. Suess PS, Tsai MY, Holzknecht RA, Horowitz M, Tuchman M (1992) Screening for gene deletions and known mutations in 13 patients with ornithine transcarbamylase deficiency. Biochem Med Metab Biol 47: 250–259

  21. Tsai MY, Holzknecht RA, Tuchman M (1993) Single-strand conformational polymorphism and direct sequencing applied to carrier testing in families with ornithine transcarbamylase deficiency. Hum Genet 91: 321–325

  22. Tuchman M (1993) Mutations and polymorphisms in the human ornithine transcarbamylase gene. Hum Mutat 2: 174–178

  23. Veres G, Gibbs RA, Scherer SE, Caskey CT (1987) The molecular basis of the sparse fur mouse mutation. Science 237: 415–417

Download references

Author information

Rights and permissions

Reprints and Permissions

About this article

Cite this article

García-Pérez, M.A., Sanjurjo, P. & Rubio, V. Demonstration of the spf-ash mutation in Spanish patients with ornithine transcarbamylase deficiency of moderate severity. Hum Genet 95, 183–186 (1995). https://doi.org/10.1007/BF00209398

Download citation

Keywords

  • Restriction Site
  • Direct Sequencing
  • Donor Site
  • Ornithine
  • Rodent Model