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  • Original Paper
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Functional characterization and role of INrf2 in antioxidant response element-mediated expression and antioxidant induction of NAD(P)H:quinone oxidoreductase1 gene

Oncogene volume 20, pages 3906–3917 (2001)Cite this article

Abstract

Antioxidant response element (ARE) and nuclear transcription factor Nrf2 are known to regulate expression and coordinated induction of NQO1 and other detoxifying enzyme genes in response to antioxidants and xenobiotics. A cytosolic inhibitor of Nrf2, INrf2, that retains Nrf2 in the cytoplasm, was cloned and sequenced. Treatment of cells with antioxidants and xenobiotics results in the release of Nrf2 from INrf2. Nrf2 then moves in the nucleus. This leads to the induction of ARE-mediated NQO1 and other detoxifying enzyme genes expression. INrf2 after dissociation from Nrf2 remains in the cytosol. Overexpression of INrf2 repressed ARE-mediated NQO1 gene expression. Deletion mapping of INrf2 revealed the requirement of KELCH domain (amino acid residues 361–597) and C-terminal region (amino acid residues 598–624) in retention of Nrf2 in the cytosol. Both these regions of INrf2 independently retained Nrf2 in the cytosol leading to the repression of ARE-mediated NQO1 gene expression. These results may indicate that two different regions of INrf2 interact with a single molecule of Nrf2 or two or more molecules of Nrf2 interact with a single molecule of INrf2. The transcription of Nrf2 and INrf2 did not change in response to antioxidants and xenobiotics. This indicated that INrf2 and/or Nrf2 might be post-transcriptionally modified in response to antioxidants and xenobiotics leading to the release of Nrf2 from INrf2 and induction of ARE-mediated NQO1 and other detoxifying enzyme genes expression.

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Abbreviations

NQO1:

NAD(P)H:Quinone Oxidoreductase1

EGFP:

Enhanced Green Fluorescent Protein

ARE:

Antioxidant Response Element

hARE:

human NQO1 gene ARE

t-BHQ:

tert-Butyl Hydroquinone

ROS:

Reactive Oxygen Species

QT6:

Quail Fibroblast cells

INrf2:

Inhibitor of Nrf2

References

  • Alam J, Stewart D, Touchard C, Boinapally S, Choi MK, Cook JL . 1999 J. Biol. Chem. 274: 26071–26078

  • Albagli O, Dhordain P, Deweindt C, Lecocq G, Leprince D . 1995 Cell. Growth. Differ. 6: 1193–1198

  • Bauer CE, Elsen S, Bird TH . 1999 Ann. Rev. Microbiol. 53: 495–523

  • Bergelson S, Pinkus R, Daniel V . 1994 Oncogene. 9: 565–571

  • Chan PK, Bloom DA, Hoang TT . 1999 Biochem. Biophys. Res. Commun. 264: 305–309

  • Chan JY, Han X, Kan YW . 1993 Proc. Natl. Acad. Sci. USA. 90: 11371–11375

  • Choi AM, Alam J . 1996 Am. J. Respir. Cell. Mol. Biol. 15: 9–19

  • De Long MJ, Santamaria AB, Talalay P . 1987 Carcinogenesis 8: 1549–1553

  • Dhakshinamoorthy S, Long II DJ, Jaiswal AK . 2000 Curr. Top. Cell. Regul. 36: 201–216

  • Dhakshinamoorthy S, Jaiswal AK . 2000 J. Biol. Chem. 275: 40134–40141

  • Huang HC, Nguyen T, Pickett CB . 2000 Proc. Natl. Acad. Sci. USA 97: 12475–12480

  • Hunter T, Karin M . 1992 Cell 70: 375–387

  • Itoh K, Wakabayashi N, Katoh Y, Ishii T, Igarashi K, Engel JD, Yamamoto M . 1999 Genes Dev. 13: 76–86

  • Jaiswal AK, Burnett P, Adesnik M, McBride OW . 1990 Biochemistry 29: 1899–1906

  • Jaiswal AK . 1994 J. Biol. Chem. 269: 14502–14508

  • Jaiswal AK . 1999 Molecular Medicine: Novel Findings of Gene Diagnosis, Regulation of Gene Expression, and Gene Therapy. Hashimoto I, Tsuchida S, Shinkawa H, Yagihashi S, Suda T and Uitto J (eds) pp. 103–112

  • Kim IF, Mohammadi E, Huang RCC . 1999 Gene. 228: 73–83

  • Kobayashi A, Ito E, Toki T, Kogame K, Takahashi S, Igarashi K, Hayashi N, Yamamoto MM . 1999 J. Biol. Chem. 274: 6443–6452

  • Li Y, Jaiswal AK . 1992 J. Biol. Chem. 267: 15097–15104

  • Li Y, Jaiswal AK . 1994 Eur. J. Biochem. 226: 31–39

  • Long II DJ, Waikel RL, Wang XJ, Perlaky L, Roop DR, Jaiswal AK . 2000 Cancer Res. 60: 5913–5915

  • Marini MG, Chan K, Casula L, Kan YW, Cao A, Moi P . 1997 J. Biol. Chem. 272: 16490–16497

  • Moi P, Chan K, Asunis I, Cao A, Kan YW . 1994 Proc. Natl. Acad. Sci. USA 91: 9926–9930

  • Mulkahy RT, Wartman MA, Bailey HH, Gipp JJ . 1997 J. Biol. Chem. 272: 7445–7454

  • Nguyen T, Huang HC, Pickett CB . 2000 J. Biol. Chem. 275: 15466–15473

  • Puga A, Nebert DW, Carrier F . 1992 DNA and Cell Biology 11: 269–281

  • Qiagen . 2000 The QIAexpressionist, A handbook for high-level expression and purification of 6XHis-tagged proteins Qiagen Inc, Valencia, CA, USA

  • Radjendirane V, Joseph P, Jaiswal AK . 1997 Oxidative Stress and Signal Transduction. Forman HJ and Cadenas E (eds) Chapman and Hall, New York pp. 441–475

  • Radjendirane V, Jaiswal AK . 1999 Biochem. Pharmacol. 58: 597–603

  • Rushmore TH, Pickett CB . 1993 J. Biol. Chem. 268: 11475–11478

  • Sambrook J, Fritsch EF, Maniatis T . 1989 Molecular Cloning: A Laboratory Manual Cold Spring Harbor Laboratory, Cold Spring Harbor, NY

  • Schmitt J, Hess H, Stunneberg HG . 1993 Mol. Biol. Rep. 18: 223–230

  • Schulze-Osthoff K, Bauer M, Vogt M, Wesselborg S, Baeuerle PA . 1997 Oxidative Stress and Signal Transduction. Forman HJ and Cadenas E (eds) Chapman and Hall, New York pp. 239–259

  • Talalay P, Fahey JW, Holtzclaw WD, Prestera T, Zhang Y . 1995 Toxicol. Lett. 82–83: 173–179

  • Thanos D, Maniatis T . 1995 Cell. 80: 529–532

  • Venugopal R, Jaiswal AK . 1996 Proc. Natl. Acad. Sci. USA 93: 14960–14965

  • Venugopal R, Jaiswal AK . 1998 Oncogene 17: 3145–3156

  • Wasserman W, Fahl WE . 1997 Proc. Natl. Acad. Sci. USA 94: 5361–5366

  • Wild AC, Gipp JJ, Mulcahy RT . 1998 Biochem. J. 332: 373–381

  • Wild AC, Moinova HR, Mulcahy RT . 1999 J. Biol. Chem. 274: 33627–33636

  • Yu R, Mandlekar S, Lei W, Fahl WE, Tan TH, Kong ANT . 2000a J. Biol. Chem. 275: 2322–2327

  • Yu R, Chen C, Mo YY, Hebbar V, Owuor ED, Tan TH, Kong ANT . 2000b J. Biol. Chem. 275: 39907–39913

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Acknowledgements

We are grateful to Drs Jefferson Y Chan and Yuet W Kan, both from University of California, San Francisco for providing us the cDNA encoding Nrf2. This investigation was supported by NIH grant GM47466.

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  1. Department of Pharmacology, Baylor College of Medicine, One Baylor Plaza, Houston, 77030, Texas, TX, USA

    Saravanakumar Dhakshinamoorthy & Anil K Jaiswal

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  1. Saravanakumar Dhakshinamoorthy
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  2. Anil K Jaiswal
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Dhakshinamoorthy, S., Jaiswal, A. Functional characterization and role of INrf2 in antioxidant response element-mediated expression and antioxidant induction of NAD(P)H:quinone oxidoreductase1 gene. Oncogene 20, 3906–3917 (2001). https://doi.org/10.1038/sj.onc.1204506

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