Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1986 Mar 25;14(6):2497–2510. doi: 10.1093/nar/14.6.2497

A comparative study of the regulation of cytochrome P-450 and glutathione transferase gene expression in rat liver.

V N Francis, V I Dwarki, G Padmanaban
PMCID: PMC339678  PMID: 3754327

Abstract

A cDNA clone for the Ya subunit of glutathione transferase from rat liver was constructed in E. coli. The clone hybridized to Ya and Yc subunit messenger RNAs. On the basis of experiments involving cell-free translation and hybridization to the cloned probe, it was shown that prototype inducers of cytochrome P-450 such as phenobarbitone and 3-methylcholanthrene as well as inhibitors such as CoCl2 and 3-amino-1,2,4-triazole enhanced the glutathione transferase (Ya+Yc) messenger RNA contents in rat liver. A comparative study with the induction of cytochrome P-450 (b+e) by phenobarbitone revealed that the drug manifested a striking increase in the nuclear pre-messenger RNAs for the cytochrome at 12 hr, but did not significantly affect the same in the case of glutathione transferase (Ya+Yc). 3-Amino-1,2,4-triazole and CoCl2 blocked the phenobarbitone mediated increase in cytochrome P-450 (b+e) nuclear pre-messenger RNAs. These compounds did not significantly affect the glutathione transferase (Ya+Yc) nuclear pre-messenger RNA levels. The polysomal, poly (A)- containing messenger RNAs for cytochrome P-450 (b+e) increased by 12-15 fold after phenobarbitone administration, reached a maximum around 16 hr and then decreased sharply. In comparison, the increase in the case a glutathione transferase (Ya+Yc) messenger RNAs was sluggish and steady and a value of 3-4 fold was reached around 24 hr. Run-off transcription rates for cytochrome P-450 (b+e) increased by nearly 15 fold in 4 hr after phenobarbitone administration, whereas the increase for glutathione transferase (Ya+Yc) was only 2.0 fold. At 12 hr after the drug administration, the glutathione transferase (Ya+Yc) transcription rates were near normal. Administration of 3-amino-1,2,4-triazole and CoCl2 blocked the phenobarbitone-mediated increase in the transcription of cytochrome P-450 (b+e) messenger RNAs. These compounds at best had only marginal effects on the transcription of glutathione transferase (Ya+Yc) messenger RNAs. The half-life of cytochrome P-450 (b+e) messenger RNA was estimated to be 3-4 hr, whereas that for glutathione transferase (Ya+Yc) was found to be 8-9 hr. Administration of phenobarbitone enhanced the half-life of glutathione transferase (Ya+Yc) messenger RNA by nearly two fold. It is suggested that while transcription activation may play a primary role in the induction of cytochrome P-450 (b+e), the induction of glutathione transferase (Ya+Yc) may essentially involve stabilization of the messenger RNAs.

Full text

PDF
2497

Images in this article

2501Image
on p.2501
2502Image
on p.2502
2504Image
on p.2504
2504Image
on p.2504

Selected References

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

  1. Atchison M., Adesnik M. A cytochrome P-450 multigene family. Characterization of a gene activated by phenobarbital administration. J Biol Chem. 1983 Sep 25;258(18):11285–11295. [PubMed] [Google Scholar]
  2. Brawerman G. Eukaryotic messenger RNA. Annu Rev Biochem. 1974;43(0):621–642. doi: 10.1146/annurev.bi.43.070174.003201. [DOI] [PubMed] [Google Scholar]
  3. Carne T., Tipping E., Ketterer B. The binding and catalytic activities of forms of ligandin after modification of its thiol groups. Biochem J. 1979 Feb 1;177(2):433–439. doi: 10.1042/bj1770433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Goldberg A. L., Dice J. F. Intracellular protein degradation in mammalian and bacterial cells. Annu Rev Biochem. 1974;43(0):835–869. doi: 10.1146/annurev.bi.43.070174.004155. [DOI] [PubMed] [Google Scholar]
  5. Gonzalez F. J., Kasper C. B. Sequential translocation of two phenobarbital-induced polysomal messenger ribonucleic acids from the nuclear envelope to the endoplasmic reticulum. Biochemistry. 1981 Apr 14;20(8):2292–2298. doi: 10.1021/bi00511a034. [DOI] [PubMed] [Google Scholar]
  6. Guertin M., Baril P., Bartkowiak J., Anderson A., Bélanger L. Rapid suppression of alpha 1-fetoprotein gene transcription by dexamethasone in developing rat liver. Biochemistry. 1983 Aug 30;22(18):4296–4302. doi: 10.1021/bi00287a021. [DOI] [PubMed] [Google Scholar]
  7. Habig W. H., Pabst M. J., Jakoby W. B. Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. J Biol Chem. 1974 Nov 25;249(22):7130–7139. [PubMed] [Google Scholar]
  8. Hager D. A., Burgess R. R. Elution of proteins from sodium dodecyl sulfate-polyacrylamide gels, removal of sodium dodecyl sulfate, and renaturation of enzymatic activity: results with sigma subunit of Escherichia coli RNA polymerase, wheat germ DNA topoisomerase, and other enzymes. Anal Biochem. 1980 Nov 15;109(1):76–86. doi: 10.1016/0003-2697(80)90013-5. [DOI] [PubMed] [Google Scholar]
  9. Hardwick J. P., Gonzalez F. J., Kasper C. B. Transcriptional regulation of rat liver epoxide hydratase, NADPH-Cytochrome P-450 oxidoreductase, and cytochrome P-450b genes by phenobarbital. J Biol Chem. 1983 Jul 10;258(13):8081–8085. [PubMed] [Google Scholar]
  10. Hayes J. D., Strange R. C., Percy-Robb I. W. Identification of two lithocholic acid-binding proteins. Separation of ligandin from glutathione S-transferase B. Biochem J. 1979 Sep 1;181(3):699–708. doi: 10.1042/bj1810699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Jakoby W. B. The glutathione S-transferases: a group of multifunctional detoxification proteins. Adv Enzymol Relat Areas Mol Biol. 1978;46:383–414. doi: 10.1002/9780470122914.ch6. [DOI] [PubMed] [Google Scholar]
  12. Jones P. B., Galeazzi D. R., Fisher J. M., Whitlock J. P., Jr Control of cytochrome P1-450 gene expression by dioxin. Science. 1985 Mar 22;227(4693):1499–1502. doi: 10.1126/science.3856321. [DOI] [PubMed] [Google Scholar]
  13. Kadenbach B., Jarausch J., Hartmann R., Merle P. Separation of mammalian cytochrome c oxidase into 13 polypeptides by a sodium dodecyl sulfate-gel electrophoretic procedure. Anal Biochem. 1983 Mar;129(2):517–521. doi: 10.1016/0003-2697(83)90586-9. [DOI] [PubMed] [Google Scholar]
  14. Kraus J. P., Rosenberg L. E. Purification of low-abundance messenger RNAs from rat liver by polysome immunoadsorption. Proc Natl Acad Sci U S A. 1982 Jul;79(13):4015–4019. doi: 10.1073/pnas.79.13.4015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lu A. Y., West S. B. Multiplicity of mammalian microsomal cytochromes P-45. Pharmacol Rev. 1979 Dec;31(4):277–295. [PubMed] [Google Scholar]
  16. Maccecchini M. L., Rudin Y., Schatz G. Transport of proteins across the mitochondrial outer membrane. A precursor form of the cytoplasmically made intermembrane enzyme cytochrome c peroxidase. J Biol Chem. 1979 Aug 25;254(16):7468–7471. [PubMed] [Google Scholar]
  17. Nebert D. W., Eisen H. J., Hankinson O. The Ah receptor: binding specificity only for foreign chemicals? Biochem Pharmacol. 1984 Mar 15;33(6):917–924. doi: 10.1016/0006-2952(84)90446-5. [DOI] [PubMed] [Google Scholar]
  18. Palmiter R. D. Magnesium precipitation of ribonucleoprotein complexes. Expedient techniques for the isolation of undergraded polysomes and messenger ribonucleic acid. Biochemistry. 1974 Aug 13;13(17):3606–3615. doi: 10.1021/bi00714a032. [DOI] [PubMed] [Google Scholar]
  19. Pelham H. R., Jackson R. J. An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem. 1976 Aug 1;67(1):247–256. doi: 10.1111/j.1432-1033.1976.tb10656.x. [DOI] [PubMed] [Google Scholar]
  20. Phillips I. R., Shephard E. A., Mitani F., Rabin B. R. Induction by phenobarbital of the mRNA for a specific variant of rat liver microsomal cytochrome P-450. Biochem J. 1981 Jun 15;196(3):839–851. doi: 10.1042/bj1960839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Pickett C. B., Donohue A. M., Lu A. Y., Hales B. F. Rat liver glutathione S-transferase B: the functional mRNAs specific for the Ya Yc subunits are induced differentially by phenobarbital. Arch Biochem Biophys. 1982 May;215(2):539–543. doi: 10.1016/0003-9861(82)90113-8. [DOI] [PubMed] [Google Scholar]
  22. Pickett C. B., Telakowski-Hopkins C. A., Ding G. J., Argenbright L., Lu A. Y. Rat liver glutathione S-transferases. Complete nucleotide sequence of a glutathione S-transferase mRNA and the regulation of the Ya, Yb, and Yc mRNAs by 3-methylcholanthrene and phenobarbital. J Biol Chem. 1984 Apr 25;259(8):5182–5188. [PubMed] [Google Scholar]
  23. Pickett C. B., Telakowski-Hopkins C. A., Donohue A. M., Lu A. Y., Hales B. F. Differential induction of rat hepatic cytochrome P-448 and glutathione S-transferase B messenger RNAs by 3-methylcholanthrene. Biochem Biophys Res Commun. 1982 Jan 29;104(2):611–619. doi: 10.1016/0006-291x(82)90681-7. [DOI] [PubMed] [Google Scholar]
  24. Poland A., Knutson J. C. 2,3,7,8-tetrachlorodibenzo-p-dioxin and related halogenated aromatic hydrocarbons: examination of the mechanism of toxicity. Annu Rev Pharmacol Toxicol. 1982;22:517–554. doi: 10.1146/annurev.pa.22.040182.002505. [DOI] [PubMed] [Google Scholar]
  25. Ravishankar H., Padmanaban G. Regulation of cytochrome P-450 gene expression. Studies with a cloned probe. J Biol Chem. 1985 Feb 10;260(3):1588–1592. [PubMed] [Google Scholar]
  26. Rodgers J. R., Johnson M. L., Rosen J. M. Measurement of mRNA concentration and mRNA half-life as a function of hormonal treatment. Methods Enzymol. 1985;109:572–592. doi: 10.1016/0076-6879(85)09116-9. [DOI] [PubMed] [Google Scholar]
  27. Ryan D. E., Thomas P. E., Levin W. Purification of characterization of a minor form of hepatic microsomal cytochrome P-450 from rats treated with polychlorinated biphenyls. Arch Biochem Biophys. 1982 Jun;216(1):272–288. doi: 10.1016/0003-9861(82)90212-0. [DOI] [PubMed] [Google Scholar]
  28. Thomas P. E., Reik L. M., Ryan D. E., Levin W. Induction of two immunochemically related rat liver cytochrome P-450 isozymes, cytochromes P-450c and P-450d, by structurally diverse xenobiotics. J Biol Chem. 1983 Apr 10;258(7):4590–4598. [PubMed] [Google Scholar]
  29. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tu C. P., Lai H. C., Li N. Q., Weiss M. J., Reddy C. C. The Yc and Ya subunits of rat liver glutathione S-transferases are the products of separate genes. J Biol Chem. 1984 Aug 10;259(15):9434–9439. [PubMed] [Google Scholar]
  31. Tu C. P., Weiss M. J., Reddy C. C. Subunit composition of rat liver glutathione S-transferases. Biochem Biophys Res Commun. 1982 Sep 30;108(2):461–467. doi: 10.1016/0006-291x(82)90851-8. [DOI] [PubMed] [Google Scholar]
  32. Tukey R. H., Hannah R. R., Negishi M., Nebert D. W., Eisen H. J. The Ah locus: correlation of intranuclear appearance of inducer-receptor complex with induction of cytochrome P1-450 mRNA. Cell. 1982 Nov;31(1):275–284. doi: 10.1016/0092-8674(82)90427-5. [DOI] [PubMed] [Google Scholar]
  33. Yuan P. M., Ryan D. E., Levin W., Shively J. E. Identification and localization of amino acid substitutions between two phenobarbital-inducible rat hepatic microsomal cytochromes P-450 by micro sequence analyses. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1169–1173. doi: 10.1073/pnas.80.5.1169. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES