Cytochrome P450 RNA—Protein Interactions

  • Matti A. Lang
  • Françoise Raffalli-Mathieu
Part of the Endocrine Updates book series (ENDO, volume 16)


Humans are constantly exposed to xenobiotics present in the environment, the food, or from clinical use. To counteract the toxic accumulation of foreign molecules, living organisms have evolved a defense system comprising a family of detoxifying enzymes (cytochromes P450, CYPs). Numerous xenobiotics induce CYP genes: the induction is an essential adaptive mechanism that allows organisms to adjust their detoxification capacity according to the needs. Only in a few cases are the induction mechanisms understood to some extent. While it is well admitted that both transcriptional and post-transcriptional (mRNA processing and turnover) control is important for cyp genes expression, very little is known about the post-transcriptional mechanisms Of regulation. We have investigated the post-transcriptional regulation of two CYP genes, CYP1A2 and CYP2A5, and identified RNA-binding proteins interacting with their mRNA in an inducer-dependent manner. The strategy used to characterize the RNA protein interaction and to identify the relevant RNA sequences as well as the regulatory factors are described in this chapter.


Electrophoretic Mobility Shift Assay CYP2A5 mRNA Posttranscriptional Regulation Homeostatic Regulation Heterogeneous Nuclear Ribonucleoprotein 
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  1. 1.
    Nelson DR, Koymans L, Kamataki T, Stegeman JJ, Feyereisen R, Waxman DJ, Waterman MR, Gotoh O, CoonMJ, Estabrook RW, Gunsalus IC, Nebert DW. 1996 P450 superfamily: update on new sequences, gene mapping, accession numbers and nomenclature. Pharmacogenetics. 6 (1): 1–42Google Scholar
  2. 2.
    Rendic S, Di Carlo FJ. 1997 Human cytochrome P450 enzymes: a status report summarizing their reactions, substrates, inducers, and inhibitors. Drug Metab Rev. 29 (12): 413–580.PubMedCrossRefGoogle Scholar
  3. 3.
    Porter TD, Coon MJ. 1991 Cytochrome P-450. Multiplicity of isoforms, substrates, and catalytic and regulatory mechanisms. J Biol Chem. 266 (21): 13469–72PubMedGoogle Scholar
  4. 4.
    Waxman DJ. 1999 P450 gene induction by structurally diverse xenochemicals: central role of nuclear receptors CAR, PXR, and PPAR. Arch Biochem Biophys. 369 (1): 11–23PubMedCrossRefGoogle Scholar
  5. 5.
    Zubiaga AM, Belasco JG, Greenberg ME. 1995 The nonamer UUAUUUAUU is the key AU-rich sequence motif that mediates mRNA degradation. Mol Cell Biol. 15(4): 221930.Google Scholar
  6. 6.
    SiomiH, Dreyfuss G. 1997 RNA-binding proteins as regulators of gene expression. Curr Opin Genet Dev. 7(3):345–53. Review.Google Scholar
  7. 7.
    Krecic AM, Swanson MS. 1999 hnRNP complexes: composition, structure, and function. Curr Opin Cell Biol. 11(3):363–71Google Scholar
  8. 8.
    Silver G, Krauter KS. 1990 Aryl hydrocarbon induction of rat cytochrome P-450d results from increased precursor RNA processing. Mol Cell Biol. 10 (12): 6765–8.PubMedGoogle Scholar
  9. 9.
    Aida K, Negishi M. 1991 Posttranscriptional regulation of coumarin 7-hydroxylase induction by xenobiotics in mouse liver: mRNA stabilization by pyrazole. Biochemistry. 30 (32): 8041–5.PubMedCrossRefGoogle Scholar
  10. 10.
    Kimura S, Gonzalez FJ, Nebert DW. 1984 Mouse cytochrome P3–450: complete cDNA and amino acid sequence. Nucleic Acids Res. 12 (6): 2917–28PubMedCrossRefGoogle Scholar
  11. 11.
    Geneste O, Raffalli F, Lang MA. 1996 Identification and characterization of a 44 kDa protein that binds specifically to the 3’-untranslated region of CYP2a5 mRNA: inducibility, subcellular distribution and possible role in mRNA stabilization. Biochem J. 313 (Pt 3): 1029–37.PubMedGoogle Scholar
  12. 12.
    Dignam JD, Lebovitz RM, Roeder RG. 1983 Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res. 11 (5): 1475–89.PubMedCrossRefGoogle Scholar
  13. 13.
    Gorski K, Carneiro M, Schibler U. 1986 Tissue-specific in vitro transcription from the mouse albumin promoter. Cell. 47 (5): 767–76.PubMedCrossRefGoogle Scholar
  14. 14.
    Brewer G, Ross J. 1988 Poly(A) shortening and degradation of the 3’ A+U-rich sequences of human c-myc mRNA in a cell-free system. Mol Cell Biol. 8 (4): 1697–708.PubMedGoogle Scholar
  15. 15.
    Raffalli-Mathieu F, Geneste O, Lang MA. 1997 Characterization of two nuclear proteins that interact with cytochrome P-450 1A2 mRNA. Regulation of RNA binding and possible role in the expression of the CYP1a2 gene. Eur J Biochem. 245 (1): 17–24.PubMedCrossRefGoogle Scholar
  16. 16.
    Silver G, Reid LM, Krauter KS. 1990 Dexamethasone-mediated regulation of 3methylcholanthrene-induced cytochrome P-450d mRNA accumulation in primary rat hepatocyte cultures. J Biol Chem. 265 (6): 3134–8.PubMedGoogle Scholar
  17. 17.
    Adams NH, Levi PE, Hodgson E. 1993 Regulation of cytochrome P-450 isozymes by methylenedioxyphenyl compounds. Chem Biol Interact. 86 (3): 255–74.PubMedCrossRefGoogle Scholar
  18. 18.
    Burd CG, Dreyfuss G. 1994 RNA binding specificity of hnRNP Al: significance of hnRNP Al high-affinity binding sites in pre-mRNA splicing. EMBO J. 13 (5): 1197–204.PubMedGoogle Scholar
  19. 19.
    Tilloy-Ellul A, Raffalli-Mathieu F, Lang MA. 1999 Analysis of RNA-protein interactions of mouse liver cytochrome P4502A5 mRNA. Biochem J. 339 (Pt 3): 695–703.PubMedCrossRefGoogle Scholar
  20. 20.
    Hamilton BJ, Nagy E, Malter JS, Arrick BA, Rigby WF. 1993 Association of heterogeneous nuclear ribonucleoprotein A 1 and C proteins with reiterated AUUUA sequences. J Biol Chem. 268 (12): 8881–7.PubMedGoogle Scholar
  21. 21.
    Choi YD, Grabowski PJ, Sharp PA, Dreyfuss G. 1986 Heterogeneous nuclear ribonucleoproteins: role in RNA splicing. Science 231 (4745): 1534–9.PubMedCrossRefGoogle Scholar
  22. 22.
    Pasco DS, Boyum KW, Merchant SN, Chalberg SC, Fagan JB. 1988 Transcriptional and post-transcriptional regulation of the genes encoding cytochromes P-450c and P-450d in vivo and in primary hepatocyte cultures. J Biol Chem. 263 (18): 8671–6.Google Scholar
  23. 23.
    Kimura S, Gonzalez FJ, Nebert DW. 1986 Tissue-specific expression of the mouse dioxin-inducible P(1)450 and P(3)450 genes: differential transcriptional activation and mRNA stability in liver and extrahepatic tissues. Mol Cell Biol. 6 (5): 1471–7.PubMedGoogle Scholar
  24. 24.
    Simmons DL, McQuiddy P, Kasper CB. 1987 Induction of the hepatic mixed-function oxidase system by synthetic glucocorticoids. Transcriptional and post-transcriptional regulation. J Biol Chem. 262 (1): 326–32.PubMedGoogle Scholar
  25. 25.
    Dalet C, Blanchard JM, Guzelian P, Barwick J, Hartle H, Maurel P. 1986 Cloning of a cDNA coding for P-450 LM3c from rabbit liver microsomes and regulation of its expression. Nucleic Acids Res. 14 (15): 5999–6015.PubMedCrossRefGoogle Scholar
  26. 26.
    Song BJ, Matsunaga T, Hardwick JP, Park SS, Veech RL, Yang CS, Gelboin HV, Gonzalez FI. 1987 Stabilization of cytochrome P450j messenger ribonucleic acid in the diabetic rat. Mol Endocrinol. 1 (8): 542–7.PubMedCrossRefGoogle Scholar
  27. 27.
    Roy NK, Kreamer GL, Konkle B, Grunwald C, Wirgin I. C1995 haracterization and prevalence of a polymorphism in the 3’ untranslated region of cytochrome P4501Alin cancer-prone Atlantic tomcod. Arch Biochem Biophys. 322(1):204–13.Google Scholar
  28. 28.
    Peng HM, Coon MJ. 1998 Regulation of rabbit cytochrome P450 2E1 expression in HepG2 cells by insulin and thyroid hormone. Mol Pharmacol. 54 (4): 740–7.PubMedGoogle Scholar
  29. 29.
    Kocarek TA, Zangar RC, Novak RF. 2000 Post-transcriptional regulation of rat CYP2E1 expression: role of CYP2E1 mRNA untranslated regions incontrol of translational efficiency and message stability. Arch Biochem Biophys. 376 (1): 180–90.PubMedCrossRefGoogle Scholar
  30. 30.
    Agellon LB, Cheema SK. 1997 The 3’-untranslated region of the mouse cholesterol 7alpha-hydroxylase mRNA contains elements responsive to post-transcriptional regulation by bile acids. Biochem J. 328 (Pt 2): 393–9.PubMedGoogle Scholar
  31. 31.
    Baker DM, Wang SL, Bell DJ, Drevon CA, Davis RA. 2000 One or more labile proteins regulate the stability of chimeric mRNAs containing the 3’-untranslated region of cholesterol-7alpha -hydroxylase mRNA. J Biol Chem. 275 (26): 19985–91.PubMedCrossRefGoogle Scholar
  32. 32.
    Ladomery M. 1997 Multifunctional proteins suggest connections between transcriptional and post-transcriptional processes. Bioessays 19 (10): 903–9.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2002

Authors and Affiliations

  • Matti A. Lang
    • 1
  • Françoise Raffalli-Mathieu
    • 1
  1. 1.University of UppsalaUppsalaSweden

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