Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1971 Jul;68(7):1516–1519. doi: 10.1073/pnas.68.7.1516

Hypoxanthine-Guanine Phosphoribosyltransferase Deficiency: Chemical Agents Selective for Mutant or Normal Cultured Fibroblasts in Mixed and Heterozygote Cultures

Wilfred Y Fujimoto 1,*, John H Subak-Sharpe 1,†, J Edwin Seegmiller 1,‡
PMCID: PMC389230  PMID: 5283941

Abstract

Cultured fibroblasts established from skin biopsies from patients with the Lesch-Nyhan syndrome are deficient in hypoxanthine-guanine phosphoribosyl-transferase (EC 2.4.2.8) activity. This deficiency makes possible the use of chemicals that select either for or against deficient variants in cultured fibroblasts. Two-way selection has been achieved by the use of 6-thioguanine, which selects for the deficient mutant, and azaserine, which selects to some extent for the normal allele in mixed cultures, as well as in cultures from heterozygotes. Theoretical considerations predict that the phenomenon of metabolic cooperation would tend to reinforce the former and to weaken the latter type of selection, and this is in accordance with the experimental findings.

Keywords: Lesch-Nyhan syndrome, skin biopsies, HAT medium, azaserine, 6-thioguanine

Full text

PDF
1516

Selected References

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

  1. BENNETT L. L., Jr, SCHABEL F. M., Jr, SKIPPER H. E. Studies on the mode of action of azaserine. Arch Biochem Biophys. 1956 Oct;64(2):423–436. doi: 10.1016/0003-9861(56)90286-7. [DOI] [PubMed] [Google Scholar]
  2. BROCKMAN R. W., SPARKS M. C., SIMPSON M. S. A comparison of the metabolism of purines and purine analogs by susceptible and drug-resistant bacterial and neoplastic cells. Biochim Biophys Acta. 1957 Dec;26(3):671–672. doi: 10.1016/0006-3002(57)90134-8. [DOI] [PubMed] [Google Scholar]
  3. Boyle J. A., Raivio K. O., Astrin K. H., Schulman J. D., Graf M. L., Seegmiller J. E., Jacobsen C. B. Lesch-Nyhan syndrome: preventive control by prenatal diagnosis. Science. 1970 Aug 14;169(3946):688–689. doi: 10.1126/science.169.3946.688. [DOI] [PubMed] [Google Scholar]
  4. Cox R. P., Krauss M. R., Balis M. E., Dancis J. Evidence for transfer of enzyme product as the basis of metabolic cooperation between tissue culture fibroblasts of Lesch-Nyhan disease and normal cells. Proc Natl Acad Sci U S A. 1970 Nov;67(3):1573–1579. doi: 10.1073/pnas.67.3.1573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. DeMars R., Sarto G., Felix J. S., Benke P. Lesch-Nyhan mutation: prenatal detection with amniotic fluid cells. Science. 1969 Jun 13;164(3885):1303–1305. doi: 10.1126/science.164.3885.1303. [DOI] [PubMed] [Google Scholar]
  6. Felix J. S., DeMars R. Detection of females heterozygous for the Lesch-Nyhan mutation by 8-azaguanine-resistant growth of cultured fibroblasts. J Lab Clin Med. 1971 Apr;77(4):596–604. [PubMed] [Google Scholar]
  7. Fujimoto W. Y., Seegmiller J. E. Hypoxanthine-guanine phosphoribosyltransferase deficiency: activity in normal, mutant, and heterozygote-cultured human skin fibroblasts. Proc Natl Acad Sci U S A. 1970 Mar;65(3):577–584. doi: 10.1073/pnas.65.3.577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fujimoto W. Y., Seegmiller J. E., Uhlendorf B. W., Jacobson C. B. Biochemical diagnosis of an X-linked disease in utero. Lancet. 1968 Aug 31;2(7566):511–512. doi: 10.1016/s0140-6736(68)90671-5. [DOI] [PubMed] [Google Scholar]
  9. HOEFNAGEL D., ANDREW E. D., MIREAULT N. G., BERNDT W. O. HEREDITARY CHOREOATHETOSIS, SELF-MUTILATION AND HYPERURICEMIA IN YOUNG MALES. N Engl J Med. 1965 Jul 15;273:130–135. doi: 10.1056/NEJM196507152730303. [DOI] [PubMed] [Google Scholar]
  10. Henderson J. F., Kelley W. N., Rosenbloom F. M., Seegmiller J. E. Inheritance of purine phosphoribosyltransferases in man. Am J Hum Genet. 1969 Jan;21(1):61–70. [PMC free article] [PubMed] [Google Scholar]
  11. LESCH M., NYHAN W. L. A FAMILIAL DISORDER OF URIC ACID METABOLISM AND CENTRAL NERVOUS SYSTEM FUNCTION. Am J Med. 1964 Apr;36:561–570. doi: 10.1016/0002-9343(64)90104-4. [DOI] [PubMed] [Google Scholar]
  12. LEVENBERG B., MELNICK I., BUCHANAN J. M. Biosynthesis of the purines. XV. The effect of aza-L-serine and 6-diazo-5-oxo-L-norleucine on inosinic acid biosynthesis de novo. J Biol Chem. 1957 Mar;225(1):163–176. [PubMed] [Google Scholar]
  13. LITTLEFIELD J. W. THE INOSINIC ACID PYROPHOSPHORYLASE ACTIVITY OF MOUSE FIBROBLASTS PARTIALLY RESISTANT TO 8-AZAGUANINE. Proc Natl Acad Sci U S A. 1963 Sep;50:568–573. doi: 10.1073/pnas.50.3.568. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. LYON M. F. Gene action in the X-chromosome of the mouse (Mus musculus L.). Nature. 1961 Apr 22;190:372–373. doi: 10.1038/190372a0. [DOI] [PubMed] [Google Scholar]
  15. MOORE E. C., LePAGE G. A. The metabolism of 6-thioguanine in normal and neoplastic tissues. Cancer Res. 1958 Oct;18(9):1075–1083. [PubMed] [Google Scholar]
  16. Migeon B. R., Der Kaloustian V. M., Nyhan W. L., Yough W. J., Childs B. X-linked hypoxanthine-guanine phosphoribosyl transferase deficiency: heterozygote has two clonal populations. Science. 1968 Apr 26;160(3826):425–427. doi: 10.1126/science.160.3826.425. [DOI] [PubMed] [Google Scholar]
  17. Migeon B. R. X-linked hypoxanthine-guanine phosphoribosyl transferase deficiency: detection of heterozygotes by selective medium. Biochem Genet. 1970 Jun;4(3):377–383. doi: 10.1007/BF00485754. [DOI] [PubMed] [Google Scholar]
  18. STUTTS P., BROCKMAN R. W. A biochemical basis for resistance of L1210 mouse leukemia to 6-thioguanine. Biochem Pharmacol. 1963 Feb;12:97–104. doi: 10.1016/0006-2952(63)90175-8. [DOI] [PubMed] [Google Scholar]
  19. SUBAK-SHARPE H. BIOCHEMICALLY MARKED VARIANTS OF THE SYRIAN HAMSTER FIBROBLAST CELL LINE BHK21 AND ITS DERIVATIVES. Exp Cell Res. 1965 Apr;38:106–119. doi: 10.1016/0014-4827(65)90432-5. [DOI] [PubMed] [Google Scholar]
  20. Salzmann J., DeMars R., Benke P. Single-allele expression at an X-linked hyperuricemia locus in heterozygous human cells. Proc Natl Acad Sci U S A. 1968 Jun;60(2):545–552. doi: 10.1073/pnas.60.2.545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Seegmiller J. E., Rosenbloom F. M., Kelley W. N. Enzyme defect associated with a sex-linked human neurological disorder and excessive purine synthesis. Science. 1967 Mar 31;155(3770):1682–1684. doi: 10.1126/science.155.3770.1682. [DOI] [PubMed] [Google Scholar]
  22. Shapiro S. L., Sheppard G. L., Jr, Dreifuss F. E., Newcombe D. S. X-linked recessive inheritance of a syndrome of mental retardation with hyperuricemia. Proc Soc Exp Biol Med. 1966 Jun;122(2):609–611. doi: 10.3181/00379727-122-31204. [DOI] [PubMed] [Google Scholar]
  23. Subak-Sharpe H., Bürk R. R., Pitts J. D. Metabolic co-operation between biochemically marked mammalian cells in tissue culture. J Cell Sci. 1969 Mar;4(2):353–367. doi: 10.1242/jcs.4.2.353. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES