Home LiteratureArticle Details
PMID: 19273602 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Acetylation of Nrf2 by p300/CBP augments promoter-specific DNA binding of Nrf2 during the antioxidant response.

Molecular and cellular biology ·Vol. 29 ·No. 10 ·2009-05-00 ·Pages 2658-72

Sun Z, Chin YE, Zhang DD

Abstract

To maintain intracellular redox homeostasis, genes encoding many antioxidants and detoxification enzymes are transcriptionally upregulated upon deleterious oxidative stress through the cis antioxidant responsive elements (AREs) in their promoter regions. Nrf2 is the critical transcription factor responsible for ARE-dependent transcription. We and others have previously demonstrated that Nrf2 is targeted for ubiquitin-mediated degradation by Keap1 in a redox-sensitive manner through modifications of distinct cysteine residues of Keap1. Here, we report that p300/CBP directly acetylates Nrf2 in response to arsenite-induced stress. We have identified multiple acetylated lysine residues within the Nrf2 Neh1 DNA-binding domain. Combined lysine-to-arginine mutations on the acetylation sites, with no effects on Nrf2 protein stability, compromised the DNA-binding activity of Nrf2 in a promoter-specific manner. These findings demonstrated that acetylation of Nrf2 by p300/CBP augments promoter-specific DNA binding of Nrf2 and established acetylation as a novel regulatory mechanism that functions in concert with Keap1-mediated ubiquitination in modulating the Nrf2-dependent antioxidant response.

MeSH Terms
Acetylation Amino Acid Sequence Animals Antioxidants/metabolism Cell Line Gene Expression Regulation Genes, Reporter Humans Intracellular Signaling Peptides and Proteins/genetics,metabolism Kelch-Like ECH-Associated Protein 1 Molecular Sequence Data NF-E2-Related Factor 2/genetics,metabolism Promoter Regions, Genetic Protein Binding Protein Structure, Tertiary Ubiquitin/metabolism p300-CBP Transcription Factors/genetics,metabolism
Chemicals
Antioxidants Intracellular Signaling Peptides and Proteins KEAP1 protein, human Kelch-Like ECH-Associated Protein 1 NF-E2-Related Factor 2 Ubiquitin p300-CBP Transcription Factors
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Sun Zheng
Department of Pharmacology and Toxicology, University of Arizona, Tucson, AZ 85721, USA.
Chin Y Eugene
Zhang Donna D
References (74)
74 references, click to expand
  1. Stress-dependent regulation of FOXO transcription factors by the SIRT1 deacetylase.
    Science. 2004 Mar 26;303(5666):2011-5 PMID: 14976264
  2. Regulation of gene expression by reactive oxygen.
    Annu Rev Pharmacol Toxicol. 1999;39:67-101 PMID: 10331077
  3. Direct evidence that sulfhydryl groups of Keap1 are the sensors regulating induction of phase 2 enzymes that protect against carcinogens and oxidants.
    Proc Natl Acad Sci U S A. 2002 Sep 3;99(18):11908-13 PMID: 12193649
  4. Phosphorylated CREB binds specifically to the nuclear protein CBP.
    Nature. 1993 Oct 28;365(6449):855-9 PMID: 8413673
  5. Modifying specific cysteines of the electrophile-sensing human Keap1 protein is insufficient to disrupt binding to the Nrf2 domain Neh2.
    Proc Natl Acad Sci U S A. 2005 Jul 19;102(29):10070-5 PMID: 16006525
  6. Identification of a novel Nrf2-regulated antioxidant response element (ARE) in the mouse NAD(P)H:quinone oxidoreductase 1 gene: reassessment of the ARE consensus sequence.
    Biochem J. 2003 Sep 1;374(Pt 2):337-48 PMID: 12816537
  7. The interaction between FOXO and SIRT1: tipping the balance towards survival.
    Trends Cell Biol. 2004 Aug;14(8):408-12 PMID: 15308206
  8. p300/CBP-mediated p53 acetylation is commonly induced by p53-activating agents and inhibited by MDM2.
    EMBO J. 2001 Mar 15;20(6):1331-40 PMID: 11250899
  9. Keap1 represses nuclear activation of antioxidant responsive elements by Nrf2 through binding to the amino-terminal Neh2 domain.
    Genes Dev. 1999 Jan 1;13(1):76-86 PMID: 9887101
  10. Negative regulation of forkhead transcription factor AFX (Foxo4) by CBP-induced acetylation.
    Int J Mol Med. 2003 Oct;12(4):503-8 PMID: 12964026
  11. Distinct cysteine residues in Keap1 are required for Keap1-dependent ubiquitination of Nrf2 and for stabilization of Nrf2 by chemopreventive agents and oxidative stress.
    Mol Cell Biol. 2003 Nov;23(22):8137-51 PMID: 14585973
  12. Six lysine residues on c-Myc are direct substrates for acetylation by p300.
    Biochem Biophys Res Commun. 2005 Oct 14;336(1):274-80 PMID: 16126174
  13. How many transcription factors does it take to turn on the heme oxygenase-1 gene?
    Am J Respir Cell Mol Biol. 2007 Feb;36(2):166-74 PMID: 16990612
  14. Sensitivity to carcinogenesis is increased and chemoprotective efficacy of enzyme inducers is lost in nrf2 transcription factor-deficient mice.
    Proc Natl Acad Sci U S A. 2001 Mar 13;98(6):3410-5 PMID: 11248092
  15. Nuclear import and export signals in control of Nrf2.
    J Biol Chem. 2005 Aug 12;280(32):29158-68 PMID: 15901726
  16. Genetic evidence that small maf proteins are essential for the activation of antioxidant response element-dependent genes.
    Mol Cell Biol. 2005 Sep;25(18):8044-51 PMID: 16135796
  17. Keap1 regulates the oxidation-sensitive shuttling of Nrf2 into and out of the nucleus via a Crm1-dependent nuclear export mechanism.
    Mol Cell Biol. 2005 Jun;25(11):4501-13 PMID: 15899855
  18. BRG1 interacts with Nrf2 to selectively mediate HO-1 induction in response to oxidative stress.
    Mol Cell Biol. 2006 Nov;26(21):7942-52 PMID: 16923960
  19. Acetylation-dependent signal transduction for type I interferon receptor.
    Cell. 2007 Oct 5;131(1):93-105 PMID: 17923090
  20. Inhibition of carcinogenesis by dietary polyphenolic compounds.
    Annu Rev Nutr. 2001;21:381-406 PMID: 11375442
  21. Nrf2 Neh5 domain is differentially utilized in the transactivation of cytoprotective genes.
    Biochem J. 2007 Jun 15;404(3):459-66 PMID: 17313370
  22. Regulation of E2F1 activity by acetylation.
    EMBO J. 2000 Feb 15;19(4):662-71 PMID: 10675335
  23. Oxidative stress sensor Keap1 functions as an adaptor for Cul3-based E3 ligase to regulate proteasomal degradation of Nrf2.
    Mol Cell Biol. 2004 Aug;24(16):7130-9 PMID: 15282312
  24. Keap1 is a redox-regulated substrate adaptor protein for a Cul3-dependent ubiquitin ligase complex.
    Mol Cell Biol. 2004 Dec;24(24):10941-53 PMID: 15572695
  25. Tip60-dependent acetylation of p53 modulates the decision between cell-cycle arrest and apoptosis.
    Mol Cell. 2006 Dec 28;24(6):827-39 PMID: 17189186
  26. Mechanistic studies of the Nrf2-Keap1 signaling pathway.
    Drug Metab Rev. 2006;38(4):769-89 PMID: 17145701
  27. 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
  28. Redox-regulated turnover of Nrf2 is determined by at least two separate protein domains, the redox-sensitive Neh2 degron and the redox-insensitive Neh6 degron.
    J Biol Chem. 2004 Jul 23;279(30):31556-67 PMID: 15143058
  29. Oxidative stress in neurodegenerative diseases.
    Annu Rev Pharmacol Toxicol. 1996;36:83-106 PMID: 8725383
  30. Histone acetyltransferases.
    Annu Rev Biochem. 2001;70:81-120 PMID: 11395403
  31. Small Maf proteins serve as transcriptional cofactors for keratinocyte differentiation in the Keap1-Nrf2 regulatory pathway.
    Proc Natl Acad Sci U S A. 2004 Apr 27;101(17):6379-84 PMID: 15087497
  32. The transcriptional coactivators p300 and CBP are histone acetyltransferases.
    Cell. 1996 Nov 29;87(5):953-9 PMID: 8945521
  33. Oxidative stress and atherosclerosis: its relationship to growth factors, thrombus formation and therapeutic approaches.
    Thromb Haemost. 1999 Sep;82 Suppl 1:32-7 PMID: 10695483
  34. Functional characterization of transcription regulators that interact with the electrophile response element.
    Biochem Biophys Res Commun. 2001 Nov 23;289(1):212-9 PMID: 11708801
  35. Acetylation is indispensable for p53 activation.
    Cell. 2008 May 16;133(4):612-26 PMID: 18485870
  36. Nrf1 and Nrf2 positively and c-Fos and Fra1 negatively regulate the human antioxidant response element-mediated expression of NAD(P)H:quinone oxidoreductase1 gene.
    Proc Natl Acad Sci U S A. 1996 Dec 10;93(25):14960-5 PMID: 8962164
  37. Eco1-dependent cohesin acetylation during establishment of sister chromatid cohesion.
    Science. 2008 Jul 25;321(5888):563-6 PMID: 18653893
  38. Acetylation of Foxo1 alters its DNA-binding ability and sensitivity to phosphorylation.
    Proc Natl Acad Sci U S A. 2005 Aug 9;102(32):11278-83 PMID: 16076959
  39. p300 is required for orderly G1/S transition in human cancer cells.
    Oncogene. 2007 Jan 4;26(1):21-9 PMID: 16878158
  40. Ubiquitination of Keap1, a BTB-Kelch substrate adaptor protein for Cul3, targets Keap1 for degradation by a proteasome-independent pathway.
    J Biol Chem. 2005 Aug 26;280(34):30091-9 PMID: 15983046
  41. Regulation of Nrf2 transactivation domain activity. The differential effects of mitogen-activated protein kinase cascades and synergistic stimulatory effect of Raf and CREB-binding protein.
    J Biol Chem. 2004 May 28;279(22):23052-60 PMID: 15020583
  42. Regulatory mechanisms controlling gene expression mediated by the antioxidant response element.
    Annu Rev Pharmacol Toxicol. 2003;43:233-60 PMID: 12359864
  43. Up-regulation of the human gamma-glutamylcysteine synthetase regulatory subunit gene involves binding of Nrf-2 to an electrophile responsive element.
    Biochem Biophys Res Commun. 1999 Aug 11;261(3):661-8 PMID: 10441483
  44. Heme oxygenase-1 and the vascular bed: from molecular mechanisms to therapeutic opportunities.
    Antioxid Redox Signal. 2008 Oct;10(10):1767-812 PMID: 18576916
  45. The Cap'n'Collar basic leucine zipper transcription factor Nrf2 (NF-E2 p45-related factor 2) controls both constitutive and inducible expression of intestinal detoxification and glutathione biosynthetic enzymes.
    Cancer Res. 2001 Apr 15;61(8):3299-307 PMID: 11309284
  46. 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
  47. Protection against electrophile and oxidant stress by induction of the phase 2 response: fate of cysteines of the Keap1 sensor modified by inducers.
    Proc Natl Acad Sci U S A. 2004 Feb 17;101(7):2040-5 PMID: 14764894
  48. Anti-inflammatory immunotherapy for multiple sclerosis/experimental autoimmune encephalomyelitis (EAE) disease.
    Curr Med Chem. 2005;12(25):2947-62 PMID: 16378498
  49. DNA damage-dependent acetylation of p73 dictates the selective activation of apoptotic target genes.
    Mol Cell. 2002 Jan;9(1):175-86 PMID: 11804596
  50. Molecular cloning and functional analysis of the adenovirus E1A-associated 300-kD protein (p300) reveals a protein with properties of a transcriptional adaptor.
    Genes Dev. 1994 Apr 15;8(8):869-84 PMID: 7523245
  51. Regulatory mechanisms of cellular response to oxidative stress.
    Free Radic Res. 1999 Oct;31(4):319-24 PMID: 10517536
  52. Keap1 controls postinduction repression of the Nrf2-mediated antioxidant response by escorting nuclear export of Nrf2.
    Mol Cell Biol. 2007 Sep;27(18):6334-49 PMID: 17636022
  53. Keap1-dependent proteasomal degradation of transcription factor Nrf2 contributes to the negative regulation of antioxidant response element-driven gene expression.
    J Biol Chem. 2003 Jun 13;278(24):21592-600 PMID: 12682069
  54. BTB protein Keap1 targets antioxidant transcription factor Nrf2 for ubiquitination by the Cullin 3-Roc1 ligase.
    Mol Cell Biol. 2005 Jan;25(1):162-71 PMID: 15601839
  55. The structural basis of protein acetylation by the p300/CBP transcriptional coactivator.
    Nature. 2008 Feb 14;451(7180):846-50 PMID: 18273021
  56. Functional antioxidant responsive elements.
    Proc Natl Acad Sci U S A. 1997 May 13;94(10):5361-6 PMID: 9144242
  57. Acetylation of the p53 DNA-binding domain regulates apoptosis induction.
    Mol Cell. 2006 Dec 28;24(6):841-51 PMID: 17189187
  58. Activation of p53 sequence-specific DNA binding by acetylation of the p53 C-terminal domain.
    Cell. 1997 Aug 22;90(4):595-606 PMID: 9288740
  59. Specific patterns of electrophile adduction trigger Keap1 ubiquitination and Nrf2 activation.
    J Biol Chem. 2005 Sep 9;280(36):31768-75 PMID: 15985429
  60. A molecular determinant for the establishment of sister chromatid cohesion.
    Science. 2008 Jul 25;321(5888):566-9 PMID: 18653894
  61. Different electrostatic potentials define ETGE and DLG motifs as hinge and latch in oxidative stress response.
    Mol Cell Biol. 2007 Nov;27(21):7511-21 PMID: 17785452
  62. Lysine acetylation: codified crosstalk with other posttranslational modifications.
    Mol Cell. 2008 Aug 22;31(4):449-461 PMID: 18722172
  63. Heme oxygenase-1/carbon monoxide: from basic science to therapeutic applications.
    Physiol Rev. 2006 Apr;86(2):583-650 PMID: 16601269
  64. The Keap1-BTB protein is an adaptor that bridges Nrf2 to a Cul3-based E3 ligase: oxidative stress sensing by a Cul3-Keap1 ligase.
    Mol Cell Biol. 2004 Oct;24(19):8477-86 PMID: 15367669
  65. Acetylation and deacetylation of non-histone proteins.
    Gene. 2005 Dec 19;363:15-23 PMID: 16289629
  66. The role of oxidative stress in carcinogenesis.
    Annu Rev Pharmacol Toxicol. 2004;44:239-67 PMID: 14744246
  67. Acetylation of Smc3 by Eco1 is required for S phase sister chromatid cohesion in both human and yeast.
    Mol Cell. 2008 Jul 11;31(1):143-51 PMID: 18614053
  68. A novel mutation lacking the bromodomain of the transcriptional coactivator p300 in the SiHa cervical carcinoma cell line.
    Biochem Biophys Res Commun. 2001 Feb 23;281(2):569-75 PMID: 11181085
  69. Evolutionary conserved N-terminal domain of Nrf2 is essential for the Keap1-mediated degradation of the protein by proteasome.
    Arch Biochem Biophys. 2005 Jan 15;433(2):342-50 PMID: 15581590
  70. Stimulation of NF-E2 DNA binding by CREB-binding protein (CBP)-mediated acetylation.
    J Biol Chem. 2001 Apr 6;276(14):10715-21 PMID: 11154691
  71. An important function of Nrf2 in combating oxidative stress: detoxification of acetaminophen.
    Proc Natl Acad Sci U S A. 2001 Apr 10;98(8):4611-6 PMID: 11287661
  72. CLOCK-mediated acetylation of BMAL1 controls circadian function.
    Nature. 2007 Dec 13;450(7172):1086-90 PMID: 18075593
  73. Nrf2, a Cap'n'Collar transcription factor, regulates induction of the heme oxygenase-1 gene.
    J Biol Chem. 1999 Sep 10;274(37):26071-8 PMID: 10473555
  74. Activation of Nrf2 by arsenite and monomethylarsonous acid is independent of Keap1-C151: enhanced Keap1-Cul3 interaction.
    Toxicol Appl Pharmacol. 2008 Aug 1;230(3):383-9 PMID: 18417180
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
1098-5549
DOI
10.1128/MCB.01639-08
Published
2009-05-00
Epub
2009-00-09
Pages
2658-72
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC2682049
Subset
IM
Grants
NIEHS NIH HHS · P30 ES006694 · United States
NIEHS NIH HHS · R01 ES015010 · United States
NIEHS NIH HHS · ES006694 · United States
NIEHS NIH HHS · 1R01ES015010 · United States
External Links
PubMed source DOI: 10.1128/MCB.01639-08 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2