Abstract
Expression of the enzymes galactokinase, thymidine kinase, and O6-methylguanine-DNA methyltransferase is occasionally coordinately regulated in human cell lines. We have measured the activities of these three enzymes in extracts of fibroblasts from individuals with hereditary galactokinase deficiency. These cells do not express measurable galactokinase activity. The levels of O6-methylguanine-DNA methyltransferase were in the normal range in cells from three galactokinase-deficient individuals. The activity of thymidine kinase in the affected cells was in the normal range for two of the three individuals. The reduced thymidine kinase activity in the third individual reflected the extremely poor growth of the cells in culture. Immortalization of one galactokinase-deficient cell line resulted in loss of O6-methylguanine-DNA methyltransferase activity, but the galactokinase and thymidine kinase levels remained unchanged. The data indicate that the loss of galactokinase activity in these individuals is the consequence of an alteration of gene expression which does not involve coordinate silencing with the thymidine kinase and methyltransferase loci.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Antequera, F., Boyes, J., and Bird, A. (1990). High levels of de novo methylation and altered chromatin structure at CpG islands in cell lines.Cell 62503.
Baptista, J., Brivet, M., Kadhom, N., Gautier, M., and Lemonnier, A. (1989). Cytosolic thymidine kinase activity in cultured human fibroblasts from individuals with galactokinase deficiency.Biochem. Genet. 27219.
Bradford, M. M. (1976). A rapid and sensitive method for the quantitative determination of microgram quantities of protein utilizing the principle of protein dye binding.Anal. Biochem. 72248.
Bradley, W. E. C. (1983). Mutation at autosomal loci of Chinese hamster ovary cells: Involvement of a high-frequency event silencing two linked alleles.Mol. Cell. Biol. 31172.
Cleaver, J. E., and Kraemer, K. (1989). Xeroderma pigmentosum. In Scriver, C. R., Beaudet, A. L., Sly, W. S., and Valle, D. (eds.),The Metabolic Basis of Inherited Disease McGraw-Hill, New York, pp. 2949–2971.
Day, R. S., Babich, M. A., Yarosh, D. B., and Scudiero, D. A. (1987). The role of O6-methylguanine in human cell killing, sister chromatid exchange induction and mutagenesis: A review.J. Cell Sci. Suppl. 6333.
Dunn, W. C., Foote, R. S., Hand, R. E., and Mitra, S. (1986). Cell cycle-dependent modulation of O6-methylguanine-DNA methyltransferase in C3H10T1/2 cells.Carcinogenesis 7807.
Elsevier, S. M., Kucherlapati, R. S., Nichols, E. A., Creagan, R. P., Giles, R. E., Ruddle, F. H., Willecke, K., and McDougall, J. K. (1974). Assignment of the gene for galactokinase to human chromosome 17 and its regional localisation to band q21–22.Nature 251633.
Fanconi, G. (1933). Hochgradige Galaktose-Intoleranz (Galaktose-Diabetes) bei einem Kinde mit Neurofibromatosis Recklinghausen.J. Kinderheilk 1381.
Felgner, P. L., Gadek, T. R., Holm, M., Roman, R., Chan, H. W., Wenz, M., Northrop, J. P., Ringold, G. M., and Danielsen, M. (1987). Lipofection: A highly efficient, lipid-mediated DNA transfection procedure.Proc. Natl. Acad. Sci. USA 847413.
Gitzelmann, R. (1965). Deficiency of erythrocyte galactokinase in a patient with galactose diabetes.Lancet 2670.
Gitzelmann, R., Wells, H. J., and Segal, S. (1974). Galactose metabolism in a patient with hereditary galactokinase deficiency.Eur. J. Clin. Invest. 479.
Goldgar, D. E., Green, P., Parry, D. M., and Mulvihill, J. J. (1989). Multipoint linkage analysis in neurofibromatosis type 1: An international collaboration.Am. J. Hum. Genet. 446.
Gradov, A. A., Pack, S. D., Sukoyan, M. A., Rubtsov, N. B., Bochkarev, N. M., and Serov, O. L. (1985). Regional assignment of the genes for TK1, GALK, ALDC and ESD on chromosome 8 in the American mink by chromosome-mediated gene transfer.Mol. Gen. Genet. 200433.
Green, M. H. L., Karran, P., Lowe, J. E., Priestley, A., Arlett, C. F., and Mayne, L. (1990). Variation in the loss of O6-methylguanine-DNA methyltransferase during immortalization of human fibroblasts.Carcinogenesis 11185.
Karran, P., Stephenson, C., Macpherson, P., Cairns-Smith, S., and Priestley, A. (1990). Coregulation of the human O6-methylguanine-DNA methyltransferase with two unrelated genes that are closely linked.Cancer Res. 501532.
Karran, P., and Hall, J. (1988). DNA repair by the Ada protein ofE. coli. In Eckstein, F., and Lilley, D. M. J. (eds.),Nucleic Acids and Molecular Biology, Vol 2 Springer-Verlag, Berlin, pp. 188–197.
Lindahl, T., Ljungquist, S., Siegert, W., Nyberg, B., and Sperens, B. (1977). DNA N-glycosidases. Properties of uracil-DNA glycosidase fromEscherichia coli.J. Biol. Chem. 2523286–3294.
Magee, P. N. (1989). The experimental basis for the role of nitroso compounds in human cancer.Cancer Surveys 8207.
Myrnes, B., Giercksky, K. E., and Krokan, H. (1983). Interindividual variation in the activity of O6-methylguanine-DNA methyltransferase and uracil-DNA glycosylase in human organs.Carcinogenesis 41565.
Okajima, K., Yazaki, M., and Wada, Y. (1987). Thymidine kinase activity in individuals with galactokinase deficiency.Am. J. Hum. Genet. 41503.
Pickering, W. R., and Howell, R. R. (1972). Galactokinase deficiency: Clinical and biochemical findings in a new kindred.J. Pediat. 8150.
Roberts, M., Scangos, G. A., Hart, J. T., and Ruddle, F. H. (1983). Genetic control of drug resistance: Assignment of ama-1 to Chinese hamster chromosome 7, confirmation of assignment of genes coding for TK, GALK and ACP to chromosome 7, and tentative assignment of TP1 to chromosome 8.Somat. Cell Genet. 9235.
Rydberg, B., Spurr, N., and Karran, P. (1990). cDNA cloning and chromosomal assignment of the human O6-methylguanine-DNA methyltransferase.J. Biol. Chem. 2659563.
Sagher, D., Karrison, T., Schwartz, J. L., Larson, R., Meier, P., and Strauss, B. (1988). Low O6-methylguanine-DNA alkyltransferase activity in the peripheral blood lymphocytes of patients with therapy-related acute non-lymphocytic leukemia.Cancer Res. 483084.
Schoen, R. C., Cox, S. H., and Wagner, R. P. (1984). Thymidine kinase activity of cultured cells from individuals with inherited galactokinase deficiency.Am. J. Hum. Genet. 36815.
Segal, S. (1989). Disorders of galactose metabolism. In Scriver, C. R., Beaudet, A. L., Sly, W. S., and Valle, D. (eds.),The Metabolic Basis of Inherited Disease McGraw-Hill, New York, pp. 453–480.
Soni, T., Brivet, M., Moatti, N., and Lemonnier, A. (1988). The Philadelphia variant of galactokinase in human erythrocytes: Physicochemical and catalytic properties.Clin. Chim. Acta 17597.
Xu, G., O'Connell, P., Viskochil, D., Crawthon, R., Robertson, M., Culver, M., Dunn, D., Stevens, J., Gesteland, R., White, R., and Weiss, R. (1990). The neurofibromatosis Type 1 gene encodes a protein related to GAP.Cell 62599.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Stephenson, C., Brivet, M., Gautier, M. et al. Normal expression of thymidine kinase and O6-methylguanine-DNA methyltransferase in cultured fibroblasts from individuals with hereditary galactokinase deficiency. Biochem Genet 29, 135–144 (1991). https://doi.org/10.1007/BF00554207
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00554207