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. 1991 Dec;19(25):7049–7052. doi: 10.1093/nar/19.25.7049

Molecular analysis of 11 galactosemia patients.

J K Reichardt 1
PMCID: PMC332510  PMID: 1766867

Abstract

Galactosemia is a human inborn error of galactose metabolism due to deficiency of galactose-1-phosphate uridyl transferase. In this paper, I describe the molecular analysis of genomic DNA, mRNA and protein from 11 different galactosemic patients by Southern, Northern and Western blotting. The results of these experiments lead me to conclude that galactosemia is caused mostly by missense mutations. The unusual preponderance of missense mutations in galactosemia led me to investigate its cause. I demonstrate that all 9 patients I investigated have detectable residual enzyme activity (ranging from 0.7-6.9% of normal). This finding is of potential importance in addressing the long-term complications of galactosemia.

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Selected References

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  1. Andersen M. W., Williams V. P., Helmer G. R., Jr, Fried C., Popják G. Transferase-deficiency galactosemia: evidence for the lack of a transferase protein in galactosemic red cells. Arch Biochem Biophys. 1983 Apr 1;222(1):326–331. doi: 10.1016/0003-9861(83)90530-1. [DOI] [PubMed] [Google Scholar]
  2. Antonarakis S. E., Kazazian H. H., Jr The molecular basis of hemophilia A in man. Trends Genet. 1988 Aug;4(8):233–237. doi: 10.1016/0168-9525(88)90156-4. [DOI] [PubMed] [Google Scholar]
  3. Banroques J., Schapira F., Grégori C., Dreyfus J. C. Molecular studies on galactose 1 phosphate uridylyl transferase from normal and mutant subjects. An immunological approach. Ann Hum Genet. 1983 Jul;47(Pt 3):177–185. doi: 10.1111/j.1469-1809.1983.tb00986.x. [DOI] [PubMed] [Google Scholar]
  4. Benn P. A., Kelley R. I., Mellman W. J., Amer L., Boches F. S., Markus H. B., Nichols W., Hoffman B. Reversion from deficiency of galactose-1-phosphate uridylytransferase (GALT) in an SV40-transformed human fibroblast line. Somatic Cell Genet. 1981 Nov;7(6):667–682. doi: 10.1007/BF01538756. [DOI] [PubMed] [Google Scholar]
  5. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  6. Dale G. L., Popják G. Purification of normal and inactive galactosemic galactose-1-phosphate uridylyltransferase from human red cells. J Biol Chem. 1976 Feb 25;251(4):1057–1063. [PubMed] [Google Scholar]
  7. Dieckmann C. L., Tzagoloff A. Assembly of the mitochondrial membrane system. CBP6, a yeast nuclear gene necessary for synthesis of cytochrome b. J Biol Chem. 1985 Feb 10;260(3):1513–1520. [PubMed] [Google Scholar]
  8. Flach J. E., Reichardt J. K., Elsas L. J., 2nd Sequence of a cDNA encoding human galactose-1-phosphate uridyl transferase. Mol Biol Med. 1990 Aug;7(4):365–369. [PubMed] [Google Scholar]
  9. Kaufman F. R., Xu Y. K., Ng W. G., Donnell G. N. Correlation of ovarian function with galactose-1-phosphate uridyl transferase levels in galactosemia. J Pediatr. 1988 May;112(5):754–756. doi: 10.1016/s0022-3476(88)80697-8. [DOI] [PubMed] [Google Scholar]
  10. Kelley R. I., Feinberg D. M., Segal S. Galactose-1-phosphate uridyl transferase in density-fractionated erythrocytes. Studies of normal and mutant enzymes. Hum Genet. 1989 May;82(2):99–103. doi: 10.1007/BF00284037. [DOI] [PubMed] [Google Scholar]
  11. Levy H. L., Hammersen G. Newborn screening for galactosemia and other galactose metabolic defects. J Pediatr. 1978 Jun;92(6):871–877. doi: 10.1016/s0022-3476(78)80351-5. [DOI] [PubMed] [Google Scholar]
  12. Merril C. R., Geier M. R., Petricciani J. C. Bacterial virus gene expression in human cells. Nature. 1971 Oct 8;233(5319):398–400. doi: 10.1038/233398a0. [DOI] [PubMed] [Google Scholar]
  13. Nadler H. L., Chacko C. M., Rachmeler M. Interallelic complementation in hybrid cells derived from human diploid strains deficient in galactose-1-phosphate uridyl transferase activity. Proc Natl Acad Sci U S A. 1970 Oct;67(2):976–982. doi: 10.1073/pnas.67.2.976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ng W. G., Xu Y. K., Kaufman F. R., Donnell G. N. Deficit of uridine diphosphate galactose in galactosaemia. J Inherit Metab Dis. 1989;12(3):257–266. doi: 10.1007/BF01799215. [DOI] [PubMed] [Google Scholar]
  15. Orkin S. H., Kazazian H. H., Jr The mutation and polymorphism of the human beta-globin gene and its surrounding DNA. Annu Rev Genet. 1984;18:131–171. doi: 10.1146/annurev.ge.18.120184.001023. [DOI] [PubMed] [Google Scholar]
  16. Reichardt J. K., Berg P. Cloning and characterization of a cDNA encoding human galactose-1-phosphate uridyl transferase. Mol Biol Med. 1988 Apr;5(2):107–122. [PubMed] [Google Scholar]
  17. Reichardt J. K., Woo S. L. Molecular basis of galactosemia: mutations and polymorphisms in the gene encoding human galactose-1-phosphate uridylyltransferase. Proc Natl Acad Sci U S A. 1991 Apr 1;88(7):2633–2637. doi: 10.1073/pnas.88.7.2633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rogers S., Holtzapple P. G., Mellman W. J., Segal S. Characteristics of galactose-1-phosphate uridyl transferase in intestinal mucosa of normal and galactosemic humans. Metabolism. 1970 Sep;19(9):701–708. doi: 10.1016/0026-0495(70)90067-3. [DOI] [PubMed] [Google Scholar]
  19. Russell J. D., DeMars R. UDP-glucose: alpha-D-galactose-1-phosphate uridylytransferase activity in cultured human fibroblasts. Biochem Genet. 1967 Jun;1(1):11–24. doi: 10.1007/BF00487732. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Segal S., Rogers S., Holtzapple P. G. Liver galactose-1-phosphate uridyl transferase: activity in normal and galactosemic subjects. J Clin Invest. 1971 Mar;50(3):500–506. doi: 10.1172/JCI106518. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Stout J. T., Caskey C. T. The Lesch-Nyhan syndrome: clinical, molecular and genetic aspects. Trends Genet. 1988 Jun;4(6):175–178. doi: 10.1016/0168-9525(88)90024-8. [DOI] [PubMed] [Google Scholar]
  23. Tedesco T. A., Mellman W. J. Galactosemia: evidence for a structural gene mutation. Science. 1971 May 14;172(3984):727–728. doi: 10.1126/science.172.3984.727. [DOI] [PubMed] [Google Scholar]
  24. Waggoner D. D., Buist N. R., Donnell G. N. Long-term prognosis in galactosaemia: results of a survey of 350 cases. J Inherit Metab Dis. 1990;13(6):802–818. doi: 10.1007/BF01800204. [DOI] [PubMed] [Google Scholar]
  25. Williams V. P. Purification and some properties of galactose 1-phosphate uridylyltransferase from human red cells. Arch Biochem Biophys. 1978 Nov;191(1):182–191. doi: 10.1016/0003-9861(78)90080-2. [DOI] [PubMed] [Google Scholar]

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