Molecular Basis of Galactosemia: Mutations and Polymorphisms in the Gene Encoding Human Galactose-1-Phosphate Uridylyltransferase

• Published in: Proceedings of the National Academy of Sciences - PNAS 1991-04, Vol.88 (7), p.2633-2637
• Main Authors: REICHARDT, J. K. V, WOO, S. L. C
• Format: Article
• Language: English
• Subjects: Alleles; Amino Acid Sequence; Animals; Base Sequence; Biological and medical sciences; Carbohydrates (enzymatic deficiencies). Glycogenosis; Cell Line; Cell lines; Complementary DNA; COS cells; DNA; DNA - genetics; DNA - isolation & purification; Errors of metabolism; galactosemia; Galactosemias; Galactosemias - enzymology; Galactosemias - genetics; Genes; Genetic mutation; Genetic Variation; Humans; Medical sciences; Metabolic diseases; Molecular Sequence Data; Mutagenesis, Site-Directed; Mutation; Oligonucleotide Probes; Oligonucleotides; Polymerase Chain Reaction; Polymorphism, Genetic; Sequencing; Transfection; UTP-Hexose-1-Phosphate Uridylyltransferase - genetics; UTP-Hexose-1-Phosphate Uridylyltransferase - metabolism
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.88.7.2633

We describe the molecular characterization of two mutations responsible for galactosemia, an inherited disorder of galatose metabolism that causes jaundice, cataracts, and mental retardation in humans. The coding region of galactose-1-phosphate uridylyltransferase (GALT; UDPglucose:α-D-galactose-1-phosphate uridylyltransferase, EC 2.7.7.12) was amplified by the polymerase chain reaction from total cDNA of a classic galactosemic individual and was characterized by direct sequencing of the products. Two missense mutations were identified: (i) replacement of valine-44 by methionine and (ii) replacement of methionine-142 by lysine. These mutations led to a drastic reduction in GALT activity when individual mutant cDNAs were overexpressed in a mammalian cell system, although full-length protein is synthesized in this assay. The two galactosemia mutations account for 3 of the 15 galactosemia alleles analyzed. These results suggest that galactosemia is caused by a variety of mutations, which might be responsible for the observed clinical heterogeneity of this disorder. We also present the molecular characterization of two GALT polymorphisms: (i) replacement of leucine-62 by methionine and (ii) replacement of asparagine-314 by aspartate. It appears that galactosemia mutations tend to occur in regions that are highly conserved throughout evolution while the polymorphisms change variable residues.