Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1988 Jun;85(12):4214–4217. doi: 10.1073/pnas.85.12.4214

cDNA cloning, sequence, and regulation of a major female-specific and growth hormone-inducible rat liver cytochrome P-450 active in 15 beta-hydroxylation of steroid sulfates.

P G Zaphiropoulos 1, A Mode 1, A Ström 1, C Möller 1, C Fernandez 1, J A Gustafsson 1
PMCID: PMC280397  PMID: 2837761

Abstract

cDNA clones encompassing the complete coding sequence for the female-specific rat liver microsomal 15 beta-hydroxylase have been isolated and sequenced. This cytochrome P-450 species (P-450 15 beta) has a total of 490 amino acid residues, and its deduced amino acid sequence as well as direct sequencing of the amino-terminal portion of the purified protein revealed its identity with P-450 i. P-450 15 beta was found to be 66% similar to the male-specific rat liver microsomal 16 alpha-hydroxylase, P-450 16 alpha. Furthermore, P-450 15 beta is 47% similar to the major phenobarbital-inducible cytochrome P-450 in rat liver, P-450 b, while its structural similarity to P-450 c, P-450 pregnenolone-16a-carbonitrile, and P-450 lauric acid omega-hydroxylase is less than 30%. However, the developmentally regulated cytochromes P-450 f and P-450 PB-1 both have 68% similarity to P-450 15 beta. It is therefore concluded that P-450 15 beta belongs to the same gene subfamily (P450IIC) that also includes cytochromes P-450 f, PB-1, and 16 alpha. The plasma pattern of growth hormone (GH) is known to be a major determinant for the sex-specific expression of P-450 15 beta and P-450 16 alpha. Continuous infusion of GH to hypophysectomized male rats causes an increase in P-450 15 beta mRNA after 6 days of treatment, while intermittent injections have no effect. On the other hand, no effect on P-450 16 alpha mRNA is observed after continuous administration of GH, while intermittent injections cause an increase after 2 days of treatment. This relatively long period before P-450 15 beta and P-450 16 alpha mRNA induction is seen might indicate that protein factors mediating GH action are involved.

Full text

PDF
4214

Images in this article

4216Image
on p.4216
4216Image
on p.4216

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adesnik M., Atchison M. Genes for cytochrome P-450 and their regulation. CRC Crit Rev Biochem. 1986;19(3):247–305. doi: 10.3109/10409238609084657. [DOI] [PubMed] [Google Scholar]
  2. Blobel G., Potter V. R. Nuclei from rat liver: isolation method that combines purity with high yield. Science. 1966 Dec 30;154(3757):1662–1665. doi: 10.1126/science.154.3757.1662. [DOI] [PubMed] [Google Scholar]
  3. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  4. Cleveland D. W., Lopata M. A., MacDonald R. J., Cowan N. J., Rutter W. J., Kirschner M. W. Number and evolutionary conservation of alpha- and beta-tubulin and cytoplasmic beta- and gamma-actin genes using specific cloned cDNA probes. Cell. 1980 May;20(1):95–105. doi: 10.1016/0092-8674(80)90238-x. [DOI] [PubMed] [Google Scholar]
  5. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Edén S. Age- and sex-related differences in episodic growth hormone secretion in the rat. Endocrinology. 1979 Aug;105(2):555–560. doi: 10.1210/endo-105-2-555. [DOI] [PubMed] [Google Scholar]
  7. Fujii-Kuriyama Y., Mizukami Y., Kawajiri K., Sogawa K., Muramatsu M. Primary structure of a cytochrome P-450: coding nucleotide sequence of phenobarbital-inducible cytochrome P-450 cDNA from rat liver. Proc Natl Acad Sci U S A. 1982 May;79(9):2793–2797. doi: 10.1073/pnas.79.9.2793. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gonzalez F. J., Kimura S., Song B. J., Pastewka J., Gelboin H. V., Hardwick J. P. Sequence of two related P-450 mRNAs transcriptionally increased during rat development. An R.dre.1 sequence occupies the complete 3' untranslated region of a liver mRNA. J Biol Chem. 1986 Aug 15;261(23):10667–10672. [PubMed] [Google Scholar]
  9. Gonzalez F. J., Nebert D. W., Hardwick J. P., Kasper C. B. Complete cDNA and protein sequence of a pregnenolone 16 alpha-carbonitrile-induced cytochrome P-450. A representative of a new gene family. J Biol Chem. 1985 Jun 25;260(12):7435–7441. [PubMed] [Google Scholar]
  10. Haniu M., Ryan D. E., Iida S., Lieber C. S., Levin W., Shively J. E. NH2-terminal sequence analyses of four rat hepatic microsomal cytochromes P-450. Arch Biochem Biophys. 1984 Dec;235(2):304–311. doi: 10.1016/0003-9861(84)90202-9. [DOI] [PubMed] [Google Scholar]
  11. Hardwick J. P., Song B. J., Huberman E., Gonzalez F. J. Isolation, complementary DNA sequence, and regulation of rat hepatic lauric acid omega-hydroxylase (cytochrome P-450LA omega). Identification of a new cytochrome P-450 gene family. J Biol Chem. 1987 Jan 15;262(2):801–810. [PubMed] [Google Scholar]
  12. Kawajiri K., Gotoh O., Sogawa K., Tagashira Y., Muramatsu M., Fujii-Kuriyama Y. Coding nucleotide sequence of 3-methylcholanthrene-inducible cytochrome P-450d cDNA from rat liver. Proc Natl Acad Sci U S A. 1984 Mar;81(6):1649–1653. doi: 10.1073/pnas.81.6.1649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Leighton J. K., DeBrunner-Vossbrinck B. A., Kemper B. Isolation and sequence analysis of three cloned cDNAs for rabbit liver proteins that are related to rabbit cytochrome P-450 (form 2), the major phenobarbital-inducible form. Biochemistry. 1984 Jan 17;23(2):204–210. doi: 10.1021/bi00297a005. [DOI] [PubMed] [Google Scholar]
  14. MacGeoch C., Morgan E. T., Cordell B., Gustafsson J. A. Growth hormone regulates expression of rat liver cytochrome P-450 15 beta at a pretranslational level. Biochem Biophys Res Commun. 1987 Mar 13;143(2):782–788. doi: 10.1016/0006-291x(87)91422-7. [DOI] [PubMed] [Google Scholar]
  15. MacGeoch C., Morgan E. T., Gustafsson J. A. Hypothalamo-pituitary regulation of cytochrome P-450(15) beta apoprotein levels in rat liver. Endocrinology. 1985 Nov;117(5):2085–2092. doi: 10.1210/endo-117-5-2085. [DOI] [PubMed] [Google Scholar]
  16. MacGeoch C., Morgan E. T., Halpert J., Gustafsson J. A. Purification, characterization, and pituitary regulation of the sex-specific cytochrome P-450 15 beta-hydroxylase from liver microsomes of untreated female rats. J Biol Chem. 1984 Dec 25;259(24):15433–15439. [PubMed] [Google Scholar]
  17. Morgan E. T., MacGeoch C., Gustafsson J. A. Hormonal and developmental regulation of expression of the hepatic microsomal steroid 16 alpha-hydroxylase cytochrome P-450 apoprotein in the rat. J Biol Chem. 1985 Oct 5;260(22):11895–11898. [PubMed] [Google Scholar]
  18. Morgan E. T., MacGeoch C., Gustafsson J. A. Sexual differentiation of cytochrome P-450 in rat liver. Evidence for a constitutive isozyme as the male-specific 16 alpha-hydroxylase. Mol Pharmacol. 1985 Apr;27(4):471–479. [PubMed] [Google Scholar]
  19. Morohashi K., Fujii-Kuriyama Y., Okada Y., Sogawa K., Hirose T., Inayama S., Omura T. Molecular cloning and nucleotide sequence of cDNA for mRNA of mitochondrial cytochrome P-450(SCC) of bovine adrenal cortex. Proc Natl Acad Sci U S A. 1984 Aug;81(15):4647–4651. doi: 10.1073/pnas.81.15.4647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Nebert D. W., Adesnik M., Coon M. J., Estabrook R. W., Gonzalez F. J., Guengerich F. P., Gunsalus I. C., Johnson E. F., Kemper B., Levin W. The P450 gene superfamily: recommended nomenclature. DNA. 1987 Feb;6(1):1–11. doi: 10.1089/dna.1987.6.1. [DOI] [PubMed] [Google Scholar]
  21. Sanger F., Coulson A. R., Barrell B. G., Smith A. J., Roe B. A. Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing. J Mol Biol. 1980 Oct 25;143(2):161–178. doi: 10.1016/0022-2836(80)90196-5. [DOI] [PubMed] [Google Scholar]
  22. Ström A., Mode A., Morgan E., Gustafsson J. A. Pretranslational hormonal control of male-specific cytochrome P-450 16 alpha in rat liver. Biochem Soc Trans. 1987 Aug;15(4):575–576. doi: 10.1042/bst0150575. [DOI] [PubMed] [Google Scholar]
  23. Tukey R. H., Okino S., Barnes H., Griffin K. J., Johnson E. F. Multiple gene-like sequences related to the rabbit hepatic progesterone 21-hydroxylase cytochrome P-450 1. J Biol Chem. 1985 Oct 25;260(24):13347–13354. [PubMed] [Google Scholar]
  24. Yabusaki Y., Shimizu M., Murakami H., Nakamura K., Oeda K., Ohkawa H. Nucleotide sequence of a full-length cDNA coding for 3-methylcholanthrene-induced rat liver cytochrome P-450MC. Nucleic Acids Res. 1984 Mar 26;12(6):2929–2938. doi: 10.1093/nar/12.6.2929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Yamazoe Y., Shimada M., Murayama N., Kato R. Suppression of levels of phenobarbital-inducible rat liver cytochrome P-450 by pituitary hormone. J Biol Chem. 1987 May 25;262(15):7423–7428. [PubMed] [Google Scholar]
  26. Yoshioka H., Morohashi K., Sogawa K., Miyata T., Kawajiri K., Hirose T., Inayama S., Fujii-Kuriyama Y., Omura T. Structural analysis and specific expression of microsomal cytochrome P-450(M-1) mRNA in male rat livers. J Biol Chem. 1987 Feb 5;262(4):1706–1711. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES