Biomaterial Database

Structural organization of the gene for rat enoyl-CoA hydratase:3-hydroxyacyl-CoA dehydrogenase bifunctional enzyme. 1987

N Ishii, and M Hijikata, and T Osumi, and T Hashimoto

Enoyl-CoA hydratase:3-hydroxyacyl-CoA dehydrogenase bifunctional enzyme is a monomeric protein which catalyzes the second and the third reactions of the peroxisomal beta-oxidation system. We cloned the gene for this enzyme from rat genomic libraries. The gene spans about 31 kilobases and consists of seven exons and six introns. The transcription initiation site was located 24 nucleotides upstream of the initiator methionine codon, ATG, determined by S1 nuclease mapping and primer extension analysis. The 5'-flanking region of the gene lacks typical TATA and CCAAT sequences, but contains G + C-rich sequences, including one CCGCCC ("GC" box) and two related GC hexanucleotides. Some of the structural features of the 5'-flanking region of this gene are shared by the 5'-upstream sequence of the gene for acyl-CoA oxidase, which catalyzes the first reaction of the peroxisomal beta-oxidation system and is induced coordinately with the bifunctional enzyme. Southern blot analysis of rat genomic DNA indicates that the bifunctional enzyme gene occurs once per haploid genome. The amino acid sequence of the carboxyl-terminal region of the bifunctional enzyme exhibits a significant level of homology to the sequence of pig mitochondrial 3-hydroxyacyl-CoA dehydrogenase. This region of the bifunctional enzyme is encoded mainly by the last exon (Exon VII) which codes for as much as 58% of the protein sequence. Based on the data plus our unpublished findings (N. Ishii, T. Osumi, and T. Hashimoto, unpublished data), we propose that the amino-terminal domain, which is principally encoded by Exons I-V, has enoyl-CoA hydratase activity, and the carboxyl-terminal one, which is mainly coded for by Exon VII, has a 3-hydroxyacyl-CoA dehydrogenase function.

UI	MeSH Term	Description	Entries
D007535	Isomerases	A class of enzymes that catalyze geometric or structural changes within a molecule to form a single product. The reactions do not involve a net change in the concentrations of compounds other than the substrate and the product.(from Dorland, 28th ed) EC 5.	Isomerase
D009097	Multienzyme Complexes	Systems of enzymes which function sequentially by catalyzing consecutive reactions linked by common metabolic intermediates. They may involve simply a transfer of water molecules or hydrogen atoms and may be associated with large supramolecular structures such as MITOCHONDRIA or RIBOSOMES.	Complexes, Multienzyme
D009693	Nucleic Acid Hybridization	Widely used technique which exploits the ability of complementary sequences in single-stranded DNAs or RNAs to pair with each other to form a double helix. Hybridization can take place between two complimentary DNA sequences, between a single-stranded DNA and a complementary RNA, or between two RNA sequences. The technique is used to detect and isolate specific sequences, measure homology, or define other characteristics of one or both strands. (Kendrew, Encyclopedia of Molecular Biology, 1994, p503)	Genomic Hybridization,Acid Hybridization, Nucleic,Acid Hybridizations, Nucleic,Genomic Hybridizations,Hybridization, Genomic,Hybridization, Nucleic Acid,Hybridizations, Genomic,Hybridizations, Nucleic Acid,Nucleic Acid Hybridizations
D003001	Cloning, Molecular	The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells.	Molecular Cloning
D004262	DNA Restriction Enzymes	Enzymes that are part of the restriction-modification systems. They catalyze the endonucleolytic cleavage of DNA sequences which lack the species-specific methylation pattern in the host cell's DNA. Cleavage yields random or specific double-stranded fragments with terminal 5'-phosphates. The function of restriction enzymes is to destroy any foreign DNA that invades the host cell. Most have been studied in bacterial systems, but a few have been found in eukaryotic organisms. They are also used as tools for the systematic dissection and mapping of chromosomes, in the determination of base sequences of DNAs, and have made it possible to splice and recombine genes from one organism into the genome of another. EC 3.21.1.	Restriction Endonucleases,DNA Restriction Enzyme,Restriction Endonuclease,Endonuclease, Restriction,Endonucleases, Restriction,Enzymes, DNA Restriction,Restriction Enzyme, DNA,Restriction Enzymes, DNA
D004746	Enoyl-CoA Hydratase	An enzyme that catalyzes reversibly the hydration of unsaturated fatty acyl-CoA to yield beta-hydroxyacyl-CoA. It plays a role in the oxidation of fatty acids and in mitochondrial fatty acid synthesis, has broad specificity, and is most active with crotonyl-CoA. EC 4.2.1.17.	3-Hydroxyacyl CoA Hydrolyases,3-Hydroxyacyl Dehydratases,Crotonase,Enoyl Hydrase,beta-Hydroxyacyl Dehydratases,Enoyl CoA Hydratases,beta-Hydroxyacyl-CoA Dehydrases,trans-2-Enoyl-Coenzyme A Hydratase,3 Hydroxyacyl CoA Hydrolyases,3 Hydroxyacyl Dehydratases,CoA Hydratases, Enoyl,CoA Hydrolyases, 3-Hydroxyacyl,Dehydrases, beta-Hydroxyacyl-CoA,Dehydratases, 3-Hydroxyacyl,Dehydratases, beta-Hydroxyacyl,Enoyl CoA Hydratase,Hydrase, Enoyl,Hydratase, Enoyl-CoA,Hydratase, trans-2-Enoyl-Coenzyme A,Hydratases, Enoyl CoA,Hydrolyases, 3-Hydroxyacyl CoA,beta Hydroxyacyl CoA Dehydrases,beta Hydroxyacyl Dehydratases,trans 2 Enoyl Coenzyme A Hydratase
D005796	Genes	A category of nucleic acid sequences that function as units of heredity and which code for the basic instructions for the development, reproduction, and maintenance of organisms.	Cistron,Gene,Genetic Materials,Cistrons,Genetic Material,Material, Genetic,Materials, Genetic
D006836	Hydro-Lyases	Enzymes that catalyze the breakage of a carbon-oxygen bond leading to unsaturated products via the removal of water. EC 4.2.1.	Dehydratase,Dehydratases,Hydrase,Hydrases,Hydro Lyase,Hydro-Lyase,Hydro Lyases,Lyase, Hydro,Lyases, Hydro
D000595	Amino Acid Sequence	The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION.	Protein Structure, Primary,Amino Acid Sequences,Sequence, Amino Acid,Sequences, Amino Acid,Primary Protein Structure,Primary Protein Structures,Protein Structures, Primary,Structure, Primary Protein,Structures, Primary Protein
D000818	Animals	Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA.	Animal,Metazoa,Animalia