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PMID: 16426233 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Role of Nrf2 in the regulation of the Mrp2 (ABCC2) gene.

The Biochemical journal ·Vol. 395 ·No. 3 ·2006-05-01 ·Pages 599-609

Vollrath V, Wielandt AM, Iruretagoyena M, Chianale J

Abstract

The Nrf2 (nuclear factor-erythroid 2 p45-related factor 2) transcription factor regulates gene expression of the GCLC (glutamate-cysteine ligase catalytic subunit), which is a key enzyme in glutathione synthesis, and GSTs (glutathione S-transferases) via the ARE (antioxidant-response element). The Mrp2 (multidrug-resistance protein 2) pump mediates the excretion of GSH and GSSG excretion as well as endo- and xeno-biotics that are conjugated with GSH, glucuronate or sulphate. Considering that Mrp2 acts synergistically with these enzymes, we hypothesized that the regulation of Mrp2 gene expression is also dependent on Nrf2. Using BHA (butylated hydroxyanisole), which is a classical activator of the ARE-Nrf2 pathway, we observed an increase in the transcriptional activity of Mrp2, GCLC and Gsta1/Gsta2 genes in the mouse liver. A similar pattern of co-induction of Mrp2 and GCLC genes was also observed in mouse (Hepa 1-6) and human (HepG2) hepatoma cells treated with BHA, beta-NF (beta-naphthoflavone), 2,4,5-T (trichlorophenoxyacetic acid) or 2AAF (2-acetylaminofluorene), suggesting that these genes share common mechanism(s) of transcriptional activation in response to exposure to xenobiotics. To define the mechanism of Mrp2 gene induction, the 5'-flanking region of the mouse Mrp2 gene (2.0 kb) was isolated, and two ARE-like sequences were found: ARE-2 (-1391 to -1381) and ARE-1 (-95 to -85). Deletion analyses demonstrated that the proximal region (-185 to +99) contains the elements for the basal expression and xenobiotic-mediated induction of the Mrp2 gene. Gel-shift and supershift assays indicated that Nrf2-protein complexes bind ARE sequences of the Mrp2 promoter, preferentially to the ARE-1 sequence. Overexpression of Nrf2 increased ARE-1-mediated CAT (chloramphenicol acetyltransferase) gene activity, while overexpression of mutant Nrf2 protein repressed the activity. Thus Nrf2 appears to regulate Mrp2 gene expression via an ARE element located at the proximal region of its promoter in response to exposure to xenobiotics.

MeSH Terms
5' Flanking Region/genetics Animals Antioxidants/metabolism Base Sequence Bile/drug effects,metabolism Butylated Hydroxyanisole/pharmacology Catalytic Domain Cell Line, Tumor Conserved Sequence Female Gene Deletion Gene Expression Regulation/drug effects,genetics Genes, Reporter/genetics Glutamate-Cysteine Ligase/genetics,metabolism Glutathione Transferase/genetics,metabolism Humans Isoenzymes/genetics,metabolism Liver/drug effects,metabolism Mice Mitochondrial Proteins/genetics,metabolism Molecular Sequence Data Multidrug Resistance-Associated Protein 2 NF-E2-Related Factor 2/genetics,metabolism Protein Binding Response Elements/genetics Ribosomal Proteins/genetics,metabolism Saccharomyces cerevisiae Proteins/genetics,metabolism Sequence Alignment Transcriptional Activation beta-Naphthoflavone/pharmacology
Chemicals
ABCC2 protein, human Antioxidants Isoenzymes MRP2 protein, S cerevisiae Mitochondrial Proteins Multidrug Resistance-Associated Protein 2 NF-E2-Related Factor 2 Nfe2l2 protein, mouse Ribosomal Proteins Saccharomyces cerevisiae Proteins Butylated Hydroxyanisole beta-Naphthoflavone Glutathione Transferase glutathione S-transferase alpha Glutamate-Cysteine Ligase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Vollrath Valeska
Department of Gastroenterology, School of Medicine, Catholic University of Chile, Casilla 114-D, Santiago, Chile. valeska@med.puc.cl
Wielandt Ana M
Iruretagoyena Mirentxu
Chianale Jose
References (45)
45 references, click to expand
  1. The human multidrug resistance protein 2 gene: functional characterization of the 5'-flanking region and expression in hepatic cells.
    Hepatology. 1999 Dec;30(6):1507-12 PMID: 10573531
  2. The ABCs of drug transport in intestine and liver: efflux proteins limiting drug absorption and bioavailability.
    Eur J Pharm Sci. 2004 Jan;21(1):25-51 PMID: 14706810
  3. Characterization of the 5'-flanking region of the human multidrug resistance protein 2 (MRP2) gene and its regulation in comparison withthe multidrug resistance protein 3 (MRP3) gene.
    Eur J Biochem. 2000 Mar;267(5):1347-58 PMID: 10691972
  4. The Nrf2 transcription factor contributes both to the basal expression of glutathione S-transferases in mouse liver and to their induction by the chemopreventive synthetic antioxidants, butylated hydroxyanisole and ethoxyquin.
    Biochem Soc Trans. 2000 Feb;28(2):33-41 PMID: 10816095
  5. Reactive oxygen species-related induction of multidrug resistance-associated protein 2 expression in primary hepatocytes exposed to sulforaphane.
    Biochem Biophys Res Commun. 2001 Mar 23;282(1):257-63 PMID: 11264000
  6. Basal expression of the rat, but not of the human, multidrug resistance protein 2 (MRP2) gene is mediated by CBF/NF-Y and Sp1 promoter-binding sites.
    Toxicology. 2001 Oct 5;167(1):25-35 PMID: 11557127
  7. Ascorbic acid restores sensitivity to imatinib via suppression of Nrf2-dependent gene expression in the imatinib-resistant cell line.
    Exp Hematol. 2004 Apr;32(4):375-81 PMID: 15050748
  8. Co-regulation of expression of phase II metabolizing enzymes and multidrug resistance-associated protein 2.
    Ann Hepatol. 2004 Jan-Mar;3(1):11-7 PMID: 15118574
  9. Genetic dissection of systemic autoimmune disease in Nrf2-deficient mice.
    Physiol Genomics. 2004 Aug 11;18(3):261-72 PMID: 15173550
  10. Contribution of NAD(P)H:quinone oxidoreductase 1 to protection against carcinogenesis, and regulation of its gene by the Nrf2 basic-region leucine zipper and the arylhydrocarbon receptor basic helix-loop-helix transcription factors.
    Mutat Res. 2004 Nov 2;555(1-2):149-71 PMID: 15476858
  11. Determination of glutathione and glutathione disulfide in biological samples.
    Methods Enzymol. 1985;113:548-55 PMID: 4088074
  12. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.
    Anal Biochem. 1987 Apr;162(1):156-9 PMID: 2440339
  13. Xenobiotic-inducible expression of murine glutathione S-transferase Ya subunit gene is controlled by an electrophile-responsive element.
    Proc Natl Acad Sci U S A. 1990 Aug;87(16):6258-62 PMID: 2166952
  14. Transcriptional regulation of the rat NAD(P)H:quinone reductase gene. Identification of regulatory elements controlling basal level expression and inducible expression by planar aromatic compounds and phenolic antioxidants.
    J Biol Chem. 1991 Mar 5;266(7):4556-61 PMID: 1900296
  15. Catalytic and regulatory properties of the heavy subunit of rat kidney gamma-glutamylcysteine synthetase.
    J Biol Chem. 1993 Sep 15;268(26):19675-80 PMID: 8103521
  16. Modulation of gamma-glutamylcysteine synthetase large subunit mRNA expression by butylated hydroxyanisole.
    Toxicol Appl Pharmacol. 1994 May;126(1):150-5 PMID: 7910420
  17. Isolation of NF-E2-related factor 2 (Nrf2), a NF-E2-like basic leucine zipper transcriptional activator that binds to the tandem NF-E2/AP1 repeat of the beta-globin locus control region.
    Proc Natl Acad Sci U S A. 1994 Oct 11;91(21):9926-30 PMID: 7937919
  18. Fibrates induce mdr2 gene expression and biliary phospholipid secretion in the mouse.
    Biochem J. 1996 Mar 15;314 ( Pt 3):781-6 PMID: 8615769
  19. Congenital jaundice in rats with a mutation in a multidrug resistance-associated protein gene.
    Science. 1996 Feb 23;271(5252):1126-8 PMID: 8599091
  20. Constitutive and beta-naphthoflavone-induced expression of the human gamma-glutamylcysteine synthetase heavy subunit gene is regulated by a distal antioxidant response element/TRE sequence.
    J Biol Chem. 1997 Mar 14;272(11):7445-54 PMID: 9054446
  21. Overexpression of mdr2 gene by peroxisome proliferators in the mouse liver.
    J Hepatol. 1997 Jun;26(6):1331-9 PMID: 9210621
  22. An Nrf2/small Maf heterodimer mediates the induction of phase II detoxifying enzyme genes through antioxidant response elements.
    Biochem Biophys Res Commun. 1997 Jul 18;236(2):313-22 PMID: 9240432
  23. Induction of cMrp/cMoat gene expression by cisplatin, 2-acetylaminofluorene, or cycloheximide in rat hepatocytes.
    Hepatology. 1997 Oct;26(4):980-5 PMID: 9328323
  24. Databases on transcriptional regulation: TRANSFAC, TRRD and COMPEL.
    Nucleic Acids Res. 1998 Jan 1;26(1):362-7 PMID: 9399875
  25. Frequent coexpression of MRP/GS-X pump and gamma-glutamylcysteine synthetase mRNA in drug-resistant cells, untreated tumor cells, and normal mouse tissues.
    Biochem Pharmacol. 1998 Mar 1;55(5):605-15 PMID: 9515571
  26. Expression of multidrug resistance protein/GS-X pump and gamma-glutamylcysteine synthetase genes is regulated by oxidative stress.
    J Biol Chem. 1998 Nov 20;273(47):31075-85 PMID: 9813007
  27. Nrf2 and Nrf1 in association with Jun proteins regulate antioxidant response element-mediated expression and coordinated induction of genes encoding detoxifying enzymes.
    Oncogene. 1998 Dec 17;17(24):3145-56 PMID: 9872330
  28. Excretion of GSSG and glutathione conjugates mediated by MRP1 and cMOAT/MRP2.
    Semin Liver Dis. 1998;18(4):359-76 PMID: 9875554
  29. Roles of MRP2 and oatp1 in hepatocellular export of reduced glutathione.
    Semin Liver Dis. 1998;18(4):377-87 PMID: 9875555
  30. Canalicular multispecific organic anion transporter/multidrug resistance protein 2 mediates low-affinity transport of reduced glutathione.
    Biochem J. 1999 Mar 1;338 ( Pt 2):393-401 PMID: 10024515
  31. Induction of the multispecific organic anion transporter (cMoat/mrp2) gene and biliary glutathione secretion by the herbicide 2,4,5-trichlorophenoxyacetic acid in the mouse liver.
    Biochem J. 1999 Jul 1;341 ( Pt 1):105-11 PMID: 10377250
  32. Glutathione transferases.
    Annu Rev Pharmacol Toxicol. 2005;45:51-88 PMID: 15822171
  33. Molecular mechanisms of reduced glutathione transport: role of the MRP/CFTR/ABCC and OATP/SLC21A families of membrane proteins.
    Toxicol Appl Pharmacol. 2005 May 1;204(3):238-55 PMID: 15845416
  34. Induction of the multidrug resistance-associated protein family of transporters by chemical activators of receptor-mediated pathways in mouse liver.
    Drug Metab Dispos. 2005 Jul;33(7):956-62 PMID: 15833929
  35. Two domains of Nrf2 cooperatively bind CBP, a CREB binding protein, and synergistically activate transcription.
    Genes Cells. 2001 Oct;6(10):857-68 PMID: 11683914
  36. Regulation of multidrug resistance-associated protein 2 (ABCC2) by the nuclear receptors pregnane X receptor, farnesoid X-activated receptor, and constitutive androstane receptor.
    J Biol Chem. 2002 Jan 25;277(4):2908-15 PMID: 11706036
  37. Role of NRF2 in protection against hyperoxic lung injury in mice.
    Am J Respir Cell Mol Biol. 2002 Feb;26(2):175-82 PMID: 11804867
  38. Influence of redox-active compounds and PXR-activators on human MRP1 and MRP2 gene expression.
    Toxicology. 2002 Feb 28;171(2-3):137-46 PMID: 11836020
  39. Loss of the Nrf2 transcription factor causes a marked reduction in constitutive and inducible expression of the glutathione S-transferase Gsta1, Gsta2, Gstm1, Gstm2, Gstm3 and Gstm4 genes in the livers of male and female mice.
    Biochem J. 2002 Jul 15;365(Pt 2):405-16 PMID: 11991805
  40. c-Maf negatively regulates ARE-mediated detoxifying enzyme genes expression and anti-oxidant induction.
    Oncogene. 2002 Aug 8;21(34):5301-12 PMID: 12149651
  41. Regulatory mechanisms controlling gene expression mediated by the antioxidant response element.
    Annu Rev Pharmacol Toxicol. 2003;43:233-60 PMID: 12359864
  42. Modulation of gene expression by cancer chemopreventive dithiolethiones through the Keap1-Nrf2 pathway. Identification of novel gene clusters for cell survival.
    J Biol Chem. 2003 Mar 7;278(10):8135-45 PMID: 12506115
  43. Bile salt transporters: molecular characterization, function, and regulation.
    Physiol Rev. 2003 Apr;83(2):633-71 PMID: 12663868
  44. Transcription factor Nrf2 is required for the constitutive and inducible expression of multidrug resistance-associated protein 1 in mouse embryo fibroblasts.
    Biochem Biophys Res Commun. 2003 Oct 24;310(3):824-9 PMID: 14550278
  45. Conjugate export pumps of the multidrug resistance protein (MRP) family: localization, substrate specificity, and MRP2-mediated drug resistance.
    Biochim Biophys Acta. 1999 Dec 6;1461(2):377-94 PMID: 10581368
Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
1470-8728
DOI
10.1042/BJ20051518
Email
valeska@med.puc.cl
Published
2006-05-01
Pages
599-609
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1462684
Subset
IM
Databases
GENBANK
AY905402
External Links
PubMed source DOI: 10.1042/BJ20051518 PMC Full Text View journal details
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