Abstract
Recently, a subset of fucosicosis patients was identified in which the single EcoRI site in the open reading frame of the human cDNA encoding α-l-fucosidase was obliterated. We have employed the polymerase chain reaction technique to amplify α-l-fucosidase DNA from the five patients known to carry the EcoRI abnormality as well as four patients and two additional fucosidosis patients who do not carry the EcoRI abnormality. Sequence analysis of the amplified DNA has determined that the EcoRI site was destroyed by a C-T transition in the last position of the EcoRI site. This single base change results in the generation of a stop codon 120 base pairs upstream of the normal stop codon. In addition, we have determined that EcoRI cleavage of amplified DNA may be a useful diagnostic tool in the diagnosis of heterozygotes and in prenatal diagnosis of fetuses at risk for this disease.
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Alhadeff, J.A., O'Brien, J.S. (1977). Fucosidosis. Practical Enzymology of the Sphingolipidoses. R.H. Glew and S.P. Peters (eds). Alan R. Liss, Inc., New York, pp 247–281
Baserga, S.J., Benz, E.J., Jr. (1988). Nonsense mutations in the human β-globin gene affect mRNA metabolism. Proc. Natl. Acad. Sci. U.S.A. 85:2056–2060
Darby, J.K., Willems, P.J., Nakashima, P., Henke, J., Ferrell, R.E., Johnsen, J., Wijsman, E.M., Gerhard, D., Dracopoli, N.C., Housman, D., Fowler, M.L., Shows, T.B., O'Brien, J.S., Cavalli-Sforza, L.L. (1988). Restriction analysis of the structural α-l-fucosidase gene and its linkage to fucosidosis. Am. J. Hum. Genet. 43:749–755
de Wet, J.R., Fukushima, H., Dewji, N.N., Wilcox, E., O'Brien, J.S., Helinski, D.R. (1984). Chromogenic immunodetection of human serum albumin and α-l-fucosidase clones in a human hepatoma cDNA expression library. DNA 3:437–447
Durand, P., Gatti, R., Borrone, C. (1982). Fucosidosis. Genetic Errors of Glycoprotein Metabolism, P. Durand and J.S. O'Brien (eds.), Springer-Verlag, New York, pp 49–87
Fukushima, H., de Wet, J.R., O'Brien, J.S. (1985). Molecular cloning of a cDNA for human α-l-fucosidase. Proc. Natl. Acad. Sci. U.S.A. 82:1262–1265
Maniatis, T., Fritsch, E.F., Sambrook, J. (eds). (1982). Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Matsuda, I., Arashima, S., Oka, Y., Mitsuyama, T., Ariga, S., Ikeuchi, T., Ichida, T. (1975). Prenatal diagnosis of fucosidosis. Clin. Chim. Acta 63:55–60
O'Brien, J.S., Willems, P.J., Fukushima, H., de Wet J.R., Darby, J.K., Di Cioccio, R., Fowler, M.L., Shows, T.B. (1987). Molecular biology of the alpha-l-fucosidase gene and fucosidosis. Enzyme 38:45–53
Saiki, R.K., Gelfand, D.H., Stoffel, S., Scharf, S.J., Higuchi, R., Horn, G.T., Mullis, K.B., Erlich, H.A. (1988). Primer directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239:487–491
Shapiro, M.B., Senapathy, P. (1987). RNA splice junctions of different classes of eukaryotes: Sequence statistics and functional implications in gene expression. Nucleic Acids Res. 15:7155–7174
Warner, T.G., O'Brien, J.S. (1983). Genetic defects in glycoprotein metabolism. Annu. Rev. Genet. 17:395–441
Willems, P.J., Darby, J.K., Di Cioccio, R.A., Nakashima, P., Eng, C., Kretz, K.A., Cavalli-Sforza, L.L., Shooter, E.M., O'Brien, J.S. (1988) Identification of a mutation in the structural α-l-fucosidase gene in fucosidosis. Am J. Hum. Genet. 43:756–763
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This research was supported by National Institutes of Health grants HD 18983 and NS 08682 to J.S.O., GM 28428 to J.K.D. and Training Grant PHS DK 07318 to K.A.K.
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Kretz, K.A., Darby, J.K., Willems, P.J. et al. Characterization of EcoRI mutation in fucosidosis patients: A stop codon in the open reading frame. J Mol Neurosci 1, 177–180 (1989). https://doi.org/10.1007/BF02918904
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DOI: https://doi.org/10.1007/BF02918904