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PMID: 19706542 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Nitric oxide activation of Keap1/Nrf2 signaling in human colon carcinoma cells.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 106 ·No. 34 ·2009-08-25 ·Pages 14547-51

Li CQ, Kim MY, Godoy LC, Thiantanawat A, Trudel LJ, Wogan GN

Abstract

The transcription factor NF-E2-related nuclear factor 2 (Nrf2) regulates expression of genes that protect cells from oxidative damage. Here, we characterized nitric oxide (*NO)-induced Nrf2-Kelch-like ECH-associated protein 1 (Keap1) signaling and its role in counteracting *NO-induced apoptosis of human colon cancer HCT116 cells. Nrf2 was localized in the cytoplasm in control cells; *NO triggered its rapid nuclear accumulation, transcriptional activation, and up-regulation of HO-1, NQO1, and GCL, but not GST A4 and P1 subunits. Nrf2 accumulation in the nucleus was also associated with enhanced transcription and posttranscriptional modifications. (S)-nitrosation of Keap1 may contribute to nuclear accumulation of Nrf2 by facilitating its dissociation from Keap1, thus initiating *NO-mediated Nrf2-Keap1 signaling. *NO-mediated induction of ARE-dependent genes occurred well before apoptosis, as judged by caspase 3 activation. Collectively, these results show that the Nrf2-Keap1 signaling pathway mediates protective cellular responses to mitigate *NO-induced damage and may contribute to the relative resistance of HCT116 to *NO-induced cytotoxicity.

MeSH Terms
Apoptosis/drug effects Blotting, Western Caspase 3/metabolism Cell Cycle/drug effects Cell Nucleus/metabolism Cell Survival/drug effects Colonic Neoplasms/genetics,metabolism,pathology Cytoplasm/metabolism Flow Cytometry Fluorescent Antibody Technique Gene Expression Regulation, Neoplastic/drug effects HCT116 Cells Heme Oxygenase-1/genetics,metabolism Humans Intracellular Signaling Peptides and Proteins/genetics,metabolism Kelch-Like ECH-Associated Protein 1 Microscopy, Confocal NAD(P)H Dehydrogenase (Quinone)/genetics,metabolism NF-E2-Related Factor 2/genetics,metabolism Nitric Oxide/pharmacology Nitrosation Protein Transport/drug effects Reverse Transcriptase Polymerase Chain Reaction Signal Transduction/drug effects
Chemicals
Intracellular Signaling Peptides and Proteins KEAP1 protein, human Kelch-Like ECH-Associated Protein 1 NF-E2-Related Factor 2 NFE2L2 protein, human Nitric Oxide HMOX1 protein, human Heme Oxygenase-1 NAD(P)H Dehydrogenase (Quinone) NQO1 protein, human Caspase 3
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Li Chun-Qi
Department of Biological Engineering and Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Kim Min Young
Godoy Luiz C
Thiantanawat Apinya
Trudel Laura J
Wogan Gerald N
References (35)
35 references, click to expand
  1. 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
  2. Antioxidants enhance mammalian proteasome expression through the Keap1-Nrf2 signaling pathway.
    Mol Cell Biol. 2003 Dec;23(23):8786-94 PMID: 14612418
  3. Biological role of glutathione in nitric oxide-induced toxicity in cell culture and animal models.
    Free Radic Biol Med. 2005 Dec 1;39(11):1489-98 PMID: 16274884
  4. Nitric oxide delivery system for cell culture studies.
    Ann Biomed Eng. 2003 Jan;31(1):65-79 PMID: 12572657
  5. Enhanced expression of the transcription factor Nrf2 by cancer chemopreventive agents: role of antioxidant response element-like sequences in the nrf2 promoter.
    Mol Cell Biol. 2002 May;22(9):2883-92 PMID: 11940647
  6. Nitric oxide-induced transcriptional up-regulation of protective genes by Nrf2 via the antioxidant response element counteracts apoptosis of neuroblastoma cells.
    J Biol Chem. 2004 May 7;279(19):20096-107 PMID: 14985350
  7. Keap1 modification and nuclear accumulation in response to S-nitrosocysteine.
    Free Radic Biol Med. 2008 Feb 15;44(4):692-8 PMID: 18062931
  8. Specific patterns of electrophile adduction trigger Keap1 ubiquitination and Nrf2 activation.
    J Biol Chem. 2005 Sep 9;280(36):31768-75 PMID: 15985429
  9. Nrf2 signaling in coordinated activation of antioxidant gene expression.
    Free Radic Biol Med. 2004 May 15;36(10):1199-207 PMID: 15110384
  10. Nuclear import and export signals in control of Nrf2.
    J Biol Chem. 2005 Aug 12;280(32):29158-68 PMID: 15901726
  11. Nrf2 as a novel molecular target for chemoprevention.
    Cancer Lett. 2005 Jun 28;224(2):171-84 PMID: 15914268
  12. 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
  13. Protection against electrophile and oxidative stress by induction of phase 2 genes: the quest for the elusive sensor that responds to inducers.
    Adv Enzyme Regul. 2004;44:335-67 PMID: 15581500
  14. Small maf (MafG and MafK) proteins negatively regulate antioxidant response element-mediated expression and antioxidant induction of the NAD(P)H:Quinone oxidoreductase1 gene.
    J Biol Chem. 2000 Dec 22;275(51):40134-41 PMID: 11013233
  15. 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
  16. 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
  17. Degradation of transcription factor Nrf2 via the ubiquitin-proteasome pathway and stabilization by cadmium.
    J Biol Chem. 2003 Jan 24;278(4):2396-402 PMID: 12441344
  18. Pharmacologic disruption of base excision repair sensitizes mismatch repair-deficient and -proficient colon cancer cells to methylating agents.
    Clin Cancer Res. 1999 Oct;5(10):2908-17 PMID: 10537360
  19. 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
  20. Thresholds of nitric oxide-mediated toxicity in human lymphoblastoid cells.
    Chem Res Toxicol. 2003 Aug;16(8):1004-13 PMID: 12924928
  21. Nitric oxide-induced genotoxicity, mitochondrial damage, and apoptosis in human lymphoblastoid cells expressing wild-type and mutant p53.
    Proc Natl Acad Sci U S A. 2002 Aug 6;99(16):10364-9 PMID: 12136132
  22. Nitric oxide as a modulator of apoptosis.
    Cancer Lett. 2005 Aug 8;226(1):1-15 PMID: 16004928
  23. Extremely potent triterpenoid inducers of the phase 2 response: correlations of protection against oxidant and inflammatory stress.
    Proc Natl Acad Sci U S A. 2005 Mar 22;102(12):4584-9 PMID: 15767573
  24. Nitric oxide stimulates Nrf2 nuclear translocation in vascular endothelium.
    Biochem Biophys Res Commun. 2003 Aug 8;307(4):973-9 PMID: 12878207
  25. Protein S-guanylation by the biological signal 8-nitroguanosine 3',5'-cyclic monophosphate.
    Nat Chem Biol. 2007 Nov;3(11):727-35 PMID: 17906641
  26. 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
  27. Nrf2 controls constitutive and inducible expression of ARE-driven genes through a dynamic pathway involving nucleocytoplasmic shuttling by Keap1.
    J Biol Chem. 2005 Sep 16;280(37):32485-92 PMID: 16000310
  28. 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
  29. Nitric oxide-induced cellular stress and p53 activation in chronic inflammation.
    Proc Natl Acad Sci U S A. 2003 Jan 7;100(1):143-8 PMID: 12518062
  30. Genotoxicity, mitochondrial damage, and apoptosis in human lymphoblastoid cells exposed to peroxynitrite generated from SIN-1.
    Chem Res Toxicol. 2002 Apr;15(4):527-35 PMID: 11952339
  31. Threshold effects of nitric oxide-induced toxicity and cellular responses in wild-type and p53-null human lymphoblastoid cells.
    Chem Res Toxicol. 2006 Mar;19(3):399-406 PMID: 16544944
  32. Nrf2-Keap1 defines a physiologically important stress response mechanism.
    Trends Mol Med. 2004 Nov;10(11):549-57 PMID: 15519281
  33. Microarray analysis reveals an antioxidant responsive element-driven gene set involved in conferring protection from an oxidative stress-induced apoptosis in IMR-32 cells.
    J Biol Chem. 2002 Jan 4;277(1):388-94 PMID: 11687587
  34. The biotin switch method for the detection of S-nitrosylated proteins.
    Sci STKE. 2001 Jun 12;2001(86):pl1 PMID: 11752655
  35. Redox signaling: nitrosylation and related target interactions of nitric oxide.
    Cell. 1994 Sep 23;78(6):931-6 PMID: 7923362
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
1091-6490
DOI
10.1073/pnas.0907539106
Published
2009-08-25
Epub
2009-00-11
Pages
14547-51
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC2732862
Subset
IM
Grants
NCI NIH HHS · P01 CA026731 · United States
NIEHS NIH HHS · P30 ES002109 · United States
NIEHS NIH HHS · ES02109 · United States
NCI NIH HHS · 5 P01 CA26731 · United States
External Links
PubMed source DOI: 10.1073/pnas.0907539106 PMC Full Text View journal details
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