Abstract
We analyzed the mitochondrial acetoacetyl-CoA thiolase gene (T2) by Southern blotting. Fifteen unrelated healthy individuals and members of five families with 3-ketothiolase deficiency (3KTD) were analyzed. We found a TaqI polymorphism, the heterozygosity of which was calculated to be 0.5 among healthy Japanese individuals. This restriction fragment length polymorphism (RFLP) proved to be useful for detecting 3KTD patients and its obligatory carriers, at the DNA level and in two out of five 3KTD families. This polymorphism was found to be generated by the presence/ absence of a TaqI site in intron 9 of the T2 gene. With in vitro amplification of the genomic region around the TaqI site, this RFLP can be detected within 2 days.
Similar content being viewed by others
References
Daum RS, Lamm PH, Mamer OA, Scriver CR (1971) A “new” disorder of isoleucine catabolism. Lancet II:1289–1290
Daum RS, Scirver CR, Mamer OA, Delvine E, Lamm PH, Goldman H (1973) An inherited disorder of isoleucine catabolism causing accumulation of a-methylacetoacetate and α-methyl-β-hydroxybutyrate, and intermittent metabolic acidosis. Pediatr Res 7:149–160
Fukao T, Yamaguchi S, Kano M, Orii T, Fujiki Y, Osumi T, Hashimoto T (1990) Molecular cloning and sequence of the complementary DNA encoding human mitochondrial acetoetyl-coenzyme. A thiolase and study of the variant enzymes in cultured fibroblasts from patients with 3-ketothiolase deficiency. J Clin Invest 86:2086–2092
Fukao T, Yamaguchi S, Tomatsu S, Orii T, Frauendienst-Egger G, Schrod L, Osumi T, Hashimoto T (1991) Evidence for structural mutation (347Ala to Thr) in a German family with 3-ketothiolase deficiency. Biochem Biophys Res Commun 179:124–129
Fukao T, Yamaguchi S, Orii T, Schutgens RBH, Osumi T, Hashimoto T (1992) Identification of three mutant alleles of the gene for mitochondrial acetoacetyl-CoA thiolase: a complete analysis of two generations of a family with 3-ketothiolase deficiency. J Clin Invest 89:474–479
Gross-Bellard M, Oudet P, Chambon P (1973) Isolation of high-molecular-weight DNA from mammalian cells. Eur J Biochem 36:32–38
Hiyama K, Sakura N, Matsumoto T, Kuhara T (1986) Deficient beta-ketothiolase activity in leukocytes from a patient with 2-methylacetoacetic aciduria. Clin Chim Acta 155:189–194
Kano M, Fukao T, Yamaguchi S, Orii T, Osumi T, Hashimoto T (1991) Structure and expression of the human mitochondrial acetoacetyl-CoA thiolase-encoding gene. Gene 109:285–290
Masuno M, Kano M, Fukao T, Yamaguchi S, Osumi T, Hashimoto T, Takahasi E, Hori T, Orii T (1992) Chromosome mapping of the human mitochondrial acetoacetyl-coenzyme A thiolase gene to band 11q22.3–q23.1 by fluorescence in situ hybridization. Cytogenet Cell Genet 60:121–122
Nagasawa H, Yamaguchi S, Orii T, Schutgens RBH, Sweetman L, Hashimoto T (1989) Heterogeneity of defects in mitochondrial acetoacetyl-CoA thiolase biosynthesis in fibroblasts from four patients with 3-ketothiolase deficiency. Pediatr Res 26:145–149
Old JM (1986) Fetal DNA analysis. In: Davies K (ed) Human genetic disease; a practical approach. IRL Press, Oxford, p 1–17
Sambrook J, Fritsch EF, Maniatis T (eds) (1989) Molecular cloning: a labortory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
Schutgens RBH, Middleton B, Blij JF, Oorthuys JWE, Veder HA, Vulsma T, Tegelaers WHH (1982) Beta-ketothiolase deficiency in a family confirmed by in vitro enzymatic assays in fibroblasts. Eur J Pediatr 139:39–42
Yamaguchi S, Orii T, Sakura N, Miyazawa S, Hashimoto T (1988) Defect in biosynthesis of mitochondrial acetoacetyl-coenzyme A thiolase in cultured fibroblasts from a boy with 3-ketothiolase deficiency. J Clin Invest 81:813–817
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kuwahara, T., Fukao, T., Kano, M. et al. Identification of TaqI polymorphism in the mitochondrial acetoacetyl-CoA thiolase gene and familial analysis of 3-ketothiolase deficiency. Hum Genet 90, 208–210 (1992). https://doi.org/10.1007/BF00220063
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00220063