Structure and expression of the human mitochondrial acetoacetyl-CoA thiolase-encoding gene

• Published in: Gene Vol. 109; no. 2; pp. 285 - 290
• Main Authors: Kano, Masatsugu, Fukao, Toshiyuki, Yamaguchi, Seiji, Orii, Tadao, Osumi, Takashi, Hashimoto, Takashi
• Format: Journal Article
• Language: English
• Published: Lausanne Elsevier 30.12.1991; Amsterdam; New York, NY
• Subjects: Acetyl-CoA C-Acetyltransferase - genetics; Amino Acid Sequence; Base Sequence; Biological and medical sciences; Blotting, Southern; Chloramphenicol O-Acetyltransferase - genetics; Chloramphenicol O-Acetyltransferase - metabolism; Cloning, Molecular; Exons - genetics; Fundamental and applied biological sciences. Psychology; genes; Genes. Genome; Humans; Introns - genetics; Mitochondria - enzymology; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; Promoter Regions, Genetic - genetics; Recombinant Proteins - biosynthesis; Restriction Mapping; Human; Nucleotide sequence; Enzyme; Acetyl-CoA acetyltransferase; Transcription promoter; Chromosome DNA; Homology; Mitochondrial DNA; Molecular cloning; Gene expression; Hybrid gene
• ISSN: 0378-1119
• DOI: 10.1016/0378-1119(91)90623-J

To examine 3-ketothiolase deficiency at the gene level, we analyzed the structure of the human mitochondrial acetoacetyl-CoA thiolase (MAT; EC 2.3.1.9)-encoding gene (MAT). From the genomic library of a normal subject in lambda EMBL3, we isolated seven overlapping clones covering the entire length of MAT and the structural organization was determined. The gene spans approx. 27 kb and contains twelve exons interrupted by eleven introns. The 5'-flanking region of the gene lacks a conventional TATA box, but is G + C-rich and contains two CAAT boxes. Included are a putative binding site for the transcription factor, Sp1, and sequences resembling the binding sites of several other transcription factors, all features characteristic of housekeeping genes. A CAT assay revealed that a 101-bp DNA fragment immediately upstream from the cap site has promoter activity, and suggested that a DNA fragment from bp -888 to -102 probably contains a negative regulatory element(s).