Cytochrome P450c17 (steroid 17α-hydroxylase/17,20 lyase): Cloning of human adrenal and testis cDNAs indicates the same gene is expressed in both tissues

B. C. Chung, J. Picado-Leonard, M. Haniu, M. Bienkowski, P. F. Hall, J. E. Shively, W. L. Miller

Research output: Contribution to journalArticle

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Abstract

P450c17 is the single enzyme mediating both 17α-hydroxylase (steroid 17α-monooxygenase, EC 1.14.99.9) and 17,20 lyase activities in the synthesis of steroid hormones. It has been suggested that different P450c17 isozymes mediate these activities in the adrenal gland and testis. We sequenced 423 of the 509 amino acids (83%) of the porcine adrenal enzyme; based on this partial sequence, a 128-fold degenerate 17-mer was synthesized and used to screen a porcine adrenal cDNA library. This yielded a 380-base cloned cDNA, which in turn was used to isolate several human adrenal cDNAs. The longest of these, λhac17-2, is 1754 base pairs long and includes the full-length coding region, the complete 3'-untranslated region, and 41 bases of the 5'-untranslated region. This cDNA encodes a protein of 508 amino acids having a predicted molecular weight of 57,379.82. High-stringency screening of a human testicular cDNA library yielded a partial clone containing 1303 identical bases. RNA gel blots and nuclease S1-protection experiments confirm that the adrenal and testicular P450c17 mRNAs are indistinguishable. These data indicate that the testis possesses a P450c17 identical to that in the adrenal. The human amino acid sequence is 66.7% homologous to the corresponding regions of the porcine sequence, and the human cDNA and amino acid sequences are 80.1 and 70.3% homologous, respectively, to bovine adrenal P450c17 cDNA. Both comparisons indicate that a central region comprising amino acid residues 160-268 is hypervariable among these species of P450c17. Comparison of the amino acid sequence of P450c17 with two other human steroidogenic cytochromes P450 show much greater homology with P450c21 (28.9%), another microsomal enzyme, than with P450scc (12.3%), a mitochondrial enzyme.

Original languageEnglish
Pages (from-to)407-411
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume84
Issue number2
DOIs
Publication statusPublished - Mar 27 1987

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Steroid 17-alpha-Hydroxylase
Cytochromes
Testis
Organism Cloning
Complementary DNA
Amino Acid Sequence
Swine
Genes
Enzymes
Gene Library
Amino Acids
5' Untranslated Regions
3' Untranslated Regions
Adrenal Glands
Base Pairing
Cytochrome P-450 Enzyme System
Isoenzymes
Clone Cells
Molecular Weight
Gels

All Science Journal Classification (ASJC) codes

  • General

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Cytochrome P450c17 (steroid 17α-hydroxylase/17,20 lyase) : Cloning of human adrenal and testis cDNAs indicates the same gene is expressed in both tissues. / Chung, B. C.; Picado-Leonard, J.; Haniu, M.; Bienkowski, M.; Hall, P. F.; Shively, J. E.; Miller, W. L.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 84, No. 2, 27.03.1987, p. 407-411.

Research output: Contribution to journalArticle

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title = "Cytochrome P450c17 (steroid 17α-hydroxylase/17,20 lyase): Cloning of human adrenal and testis cDNAs indicates the same gene is expressed in both tissues",
abstract = "P450c17 is the single enzyme mediating both 17α-hydroxylase (steroid 17α-monooxygenase, EC 1.14.99.9) and 17,20 lyase activities in the synthesis of steroid hormones. It has been suggested that different P450c17 isozymes mediate these activities in the adrenal gland and testis. We sequenced 423 of the 509 amino acids (83{\%}) of the porcine adrenal enzyme; based on this partial sequence, a 128-fold degenerate 17-mer was synthesized and used to screen a porcine adrenal cDNA library. This yielded a 380-base cloned cDNA, which in turn was used to isolate several human adrenal cDNAs. The longest of these, λhac17-2, is 1754 base pairs long and includes the full-length coding region, the complete 3'-untranslated region, and 41 bases of the 5'-untranslated region. This cDNA encodes a protein of 508 amino acids having a predicted molecular weight of 57,379.82. High-stringency screening of a human testicular cDNA library yielded a partial clone containing 1303 identical bases. RNA gel blots and nuclease S1-protection experiments confirm that the adrenal and testicular P450c17 mRNAs are indistinguishable. These data indicate that the testis possesses a P450c17 identical to that in the adrenal. The human amino acid sequence is 66.7{\%} homologous to the corresponding regions of the porcine sequence, and the human cDNA and amino acid sequences are 80.1 and 70.3{\%} homologous, respectively, to bovine adrenal P450c17 cDNA. Both comparisons indicate that a central region comprising amino acid residues 160-268 is hypervariable among these species of P450c17. Comparison of the amino acid sequence of P450c17 with two other human steroidogenic cytochromes P450 show much greater homology with P450c21 (28.9{\%}), another microsomal enzyme, than with P450scc (12.3{\%}), a mitochondrial enzyme.",
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AU - Haniu, M.

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