Structural analysis of the gene encoding human 3β-hydroxysteroid dehydrogenase/Δ5→4-isomerase

Matthew C. Lorence, C. Jo Corbin, Naohisa Kamimura, Mala S. Mahendroo, J. Ian Mason

Research output: Contribution to journalArticle

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Abstract

The structural gene encoding human 3β-hydroxysteroid dehydrogenase/Δ5→4-isomerase (3βHSD) was isolated from a human EMBL3 genomic library. The gene encompasses approximately 8 kilobases of DNA and is comprised of two large introns and three exons encoding amino acid residues 1-48, 49-103, and 104-373, respectively. The exonic sequence is identical to that of the cDNA that we previously isolated and expressed in COS 1 cells. DNA sequence analysis reveals a putative TATA (TATATAA) motif 26 basepairs up-stream of the beginning of exon I, as determined by S1 nuclease protection analysis. However, primer extension analysis using poly(A)+ RNA isolated from both placenta and corpora lutea indicates that the RNA initiates up-stream of the putative TATA motif, and that an additional 53-basepair exon, which is untranslated, is present 5′ to the first coding exon. Southern hybridization analysis of genomic DNA using a single exon probe suggests that there may be more than one copy of the gene in the human genome. In addition, we confirm from Southern analysis of genomic DNA isolated from human × hamster somatic cell hybrids that the gene is located on human chromosome 1. These findings will provide a foundation for the characterization of apparent 3βHSD clinical deficiencies when these are due to a mutation in the struc-tural gene.

Original languageEnglish (US)
Pages (from-to)1850-1855
Number of pages6
JournalMolecular Endocrinology
Volume4
Issue number12
StatePublished - 1990

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3-Hydroxysteroid Dehydrogenases
Isomerases
Exons
Genes
DNA
Nucleic Acid Hybridization
Genomic Library
Hybrid Cells
Chromosomes, Human, Pair 1
COS Cells
Corpus Luteum
Human Chromosomes
Human Genome
DNA Sequence Analysis
Cricetinae
Introns
Placenta
Complementary DNA
RNA
Amino Acids

ASJC Scopus subject areas

  • Molecular Biology
  • Endocrinology, Diabetes and Metabolism

Cite this

Structural analysis of the gene encoding human 3β-hydroxysteroid dehydrogenase/Δ5→4-isomerase. / Lorence, Matthew C.; Corbin, C. Jo; Kamimura, Naohisa; Mahendroo, Mala S.; Mason, J. Ian.

In: Molecular Endocrinology, Vol. 4, No. 12, 1990, p. 1850-1855.

Research output: Contribution to journalArticle

Lorence, MC, Corbin, CJ, Kamimura, N, Mahendroo, MS & Mason, JI 1990, 'Structural analysis of the gene encoding human 3β-hydroxysteroid dehydrogenase/Δ5→4-isomerase', Molecular Endocrinology, vol. 4, no. 12, pp. 1850-1855.
Lorence, Matthew C. ; Corbin, C. Jo ; Kamimura, Naohisa ; Mahendroo, Mala S. ; Mason, J. Ian. / Structural analysis of the gene encoding human 3β-hydroxysteroid dehydrogenase/Δ5→4-isomerase. In: Molecular Endocrinology. 1990 ; Vol. 4, No. 12. pp. 1850-1855.
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