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.
Similar content being viewed by others
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
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.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
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
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.onc.1204506
- Springer Nature Limited
Keywords
This article is cited by
-
Nrf2 in liver toxicology
Archives of Pharmacal Research (2020)
-
GSK-3β downregulates Nrf2 in cultured cortical neurons and in a rat model of cerebral ischemia-reperfusion
Scientific Reports (2016)
-
The Atypical Antipsychotic Paliperidone Regulates Endogenous Antioxidant/Anti-Inflammatory Pathways in Rat Models of Acute and Chronic Restraint Stress
Neurotherapeutics (2016)
-
The Parkinson’s-associated protein DJ-1 regulates the 20S proteasome
Nature Communications (2015)
-
Effects of dietary tert-butylhydroquinone on domoic acid metabolism and transcription of detoxification-related liver genes in red sea bream Pagrus major
Chinese Science Bulletin (2013)