Isolation and Characterization of Rat and Human cDNAs Encoding a Novel Putative Peroxisomal Enoyl-CoA Hydratase

David R. FitzPatrick, Emily Germain-Lee, David Valle

Research output: Contribution to journalArticle

Abstract

We have used a PCR-based subtractive hybridization method to identify upregulated cDNAs in the livers of rats treated with a peroxisome proliferator [clofibrate or di(2-ethylhexyl) phthalate]. After four rounds of subtractive hybridization 62 differentially hybridizing clones were partially sequenced and analyzed by sequence homology searching. Of 62, 49 were identical to 14 different upregulated rat sequences in the databank (mostly genes encoding microsomal or peroxisomal enzymes), 4 of 62 were fragments of three previously unknown genes, and 9 of 62 were false positives. Two of the unknown fragments hybridized to a single novel cDNA that was found to be more than 20-fold induced by both peroxisome proliferators. The 36-kDa predicted protein product of this cDNA shows a high degree of sequence homology to enoyl-CoA hydratases of several different species and has a C-terminal peroxisomal targeting sequence. An epitope-tagged protein product of a full-length cDNA was targeted to peroxisomes in a human cell line. We named this gene, which encodes an apparent peroxisomal enoyl-CoA hydratase, ECH1. We have also identified human ECH1 cDNA and mapped its structural gene to 19q13, 3′ to the ryanodine receptor, by hybridization to somatic cell hybrid DNA and chromosome 19-specific cosmid arrays. Possible roles for the ECH1 protein product in peroxisomal β-oxidation are discussed.

Original languageEnglish (US)
Pages (from-to)457-466
Number of pages10
JournalGenomics
Volume27
Issue number3
DOIs
StatePublished - Jun 10 1995

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Enoyl-CoA Hydratase
Complementary DNA
Peroxisome Proliferators
Sequence Homology
Genes
Chromosomes, Human, Pair 19
Clofibrate
Cosmids
Ryanodine Receptor Calcium Release Channel
Proteins
Peroxisomes
Hybrid Cells
Epitopes
Clone Cells
Databases
Cell Line
Polymerase Chain Reaction
Liver
DNA
Enzymes

ASJC Scopus subject areas

  • Genetics

Cite this

Isolation and Characterization of Rat and Human cDNAs Encoding a Novel Putative Peroxisomal Enoyl-CoA Hydratase. / FitzPatrick, David R.; Germain-Lee, Emily; Valle, David.

In: Genomics, Vol. 27, No. 3, 10.06.1995, p. 457-466.

Research output: Contribution to journalArticle

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