title: Proteomic analysis of Nrf2 deficient transgenic mice reveals cellular defence and lipid metabolism as primary Nrf2-dependent pathways in the liver.
abstract: The transcription factor Nrf2 regulates expression of multiple cellular defence proteins through the antioxidant response element (ARE). Nrf2-deficient mice (Nrf2(-/-)) are highly susceptible to xenobiotic-mediated toxicity, but the precise molecular basis of enhanced toxicity is unknown. Oligonucleotide array studies suggest that a wide range of gene products is altered constitutively, however no equivalent proteomics analyses have been conducted. To define the range of Nrf2-regulated proteins at the constitutive level, protein expression profiling of livers from Nrf2(-/-) and wild type mice was conducted using both stable isotope labelling (iTRAQ) and gel electrophoresis methods. To establish a robust reproducible list of Nrf2-dependent proteins, three independent groups of mice were analysed. Correlative network analysis (MetaCore) identified two predominant groups of Nrf2-regulated proteins. As expected, one group comprised proteins involved in phase II drug metabolism, which were down-regulated in the absence of Nrf2. Surprisingly, the most profound changes were observed amongst proteins involved in the synthesis and metabolism of fatty acids and other lipids. Importantly, we show here for the first time, that the enzyme ATP-citrate lyase, responsible for acetyl-CoA production, is negatively regulated by Nrf2. This latter finding suggests that Nrf2 is a major regulator of cellular lipid disposition in the liver.
authors.affiliation: MRC Centre for Drug Safety Science, School of Biomedical Sciences, University of Liverpool, Liverpool, Merseyside, United Kingdom. neilk@liv.ac.uk
authors.forename: Neil R
authors.initials: NR
authors.lastname: Kitteringham
authors.forename: Azman
authors.initials: A
authors.lastname: Abdullah
authors.forename: Joanne
authors.initials: J
authors.lastname: Walsh
authors.forename: Laura
authors.initials: L
authors.lastname: Randle
authors.forename: Rosalind E
authors.initials: RE
authors.lastname: Jenkins
authors.forename: Rowena
authors.initials: R
authors.lastname: Sison
authors.forename: Christopher E P
authors.initials: CE
authors.lastname: Goldring
authors.forename: Helen
authors.initials: H
authors.lastname: Powell
authors.forename: Christopher
authors.initials: C
authors.lastname: Sanderson
authors.forename: Samantha
authors.initials: S
authors.lastname: Williams
authors.forename: Larry
authors.initials: L
authors.lastname: Higgins
authors.forename: Masayuki
authors.initials: M
authors.lastname: Yamamoto
authors.forename: John
authors.initials: J
authors.lastname: Hayes
authors.forename: B Kevin
authors.initials: BK
authors.lastname: Park
mesh_terms: D001275:ATP Citrate (pro-S)-Lyase / Q000235:genetics; D000818:Animals; D000975:Antioxidants / Q000378:metabolism; D015536:Down-Regulation; D015180:Electrophoresis, Gel, Two-Dimensional; D020869:Gene Expression Profiling; D050356:Lipid Metabolism / Q000235:genetics; D008099:Liver / Q000378:metabolism; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D051267:NF-E2-Related Factor 2 / Q000172:deficiency*; D040901:Proteomics / Q000379:methods*; D015854:Up-Regulation
chemical_list: D000975:Antioxidants; D051267:NF-E2-Related Factor 2; C495636:Nfe2l2 protein, mouse; D001275:ATP Citrate (pro-S)-Lyase
references.citation: Free Radic Biol Med. 2010 Jan 15;48(2):357-71
references.pmid: 19914374
references.citation: Cancer Res. 2002 Sep 15;62(18):5196-203
references.pmid: 12234984
references.citation: J Biol Chem. 1995 Oct 13;270(41):24468-74
references.pmid: 7592662
references.citation: Nature. 1974 Nov 22;252(5481):326-8
references.pmid: 4473724
references.citation: Adv Enzyme Regul. 1993;33:281-96
references.pmid: 8356913
references.citation: J Biol Chem. 2000 Sep 8;275(36):27694-702
references.pmid: 10874044
references.citation: Proteomics. 2007 Jun;7(12):2121-9
references.pmid: 17514684
references.citation: J Biol Chem. 2000 May 26;275(21):16023-9
references.pmid: 10821856
references.citation: J Biol Chem. 1999 Sep 10;274(37):26071-8
references.pmid: 10473555
references.citation: J Biol Chem. 2000 May 19;275(20):15466-73
references.pmid: 10747902
references.citation: Proc Natl Acad Sci U S A. 1996 Nov 26;93(24):13943-8
references.pmid: 8943040
references.citation: J Biol Chem. 2001 May 25;276(21):18399-406
references.pmid: 11279008
references.citation: Electrophoresis. 2000 Apr;21(6):1037-53
references.pmid: 10786879
references.citation: Biochem Biophys Res Commun. 2001 Jan 12;280(1):286-92
references.pmid: 11162512
references.citation: Biochem J. 2002 Jul 15;365(Pt 2):405-16
references.pmid: 11991805
references.citation: Blood. 1998 May 1;91(9):3459-66
references.pmid: 9558405
references.citation: J Pharmacol Exp Ther. 2008 May;325(2):655-64
references.pmid: 18281592
references.citation: Carcinogenesis. 2009 Jun;30(6):1024-31
references.pmid: 19386581
references.citation: Methods Mol Biol. 1999;112:487-511
references.pmid: 10027273
references.citation: Nucleic Acids Res. 2003 Jul 1;31(13):3593-6
references.pmid: 12824373
references.citation: Biochem Biophys Res Commun. 1997 Jul 18;236(2):313-22
references.pmid: 9240432
references.citation: Anal Biochem. 1976 May 7;72:248-54
references.pmid: 942051
references.citation: Drug Metab Rev. 2007;39(1):235-48
references.pmid: 17364885
references.citation: Peptides. 2004 Sep;25(9):1553-63
references.pmid: 15374657
references.citation: Cancer Res. 1970 Mar;30(3):559-76
references.pmid: 4915745
references.citation: Adv Pharmacol. 1997;38:293-328
references.pmid: 8895814
references.citation: Antioxid Redox Signal. 2007 Nov;9(11):1963-70
references.pmid: 17822364
references.citation: Hepatology. 2000 Aug;32(2):321-33
references.pmid: 10915739
references.citation: Biochem J. 2003 Sep 1;374(Pt 2):337-48
references.pmid: 12816537
references.citation: Kidney Int. 2001 Oct;60(4):1343-53
references.pmid: 11576348
references.citation: Nature. 1999 Dec 16;402(6763):715-20
references.pmid: 10617183
references.citation: Toxicology. 2008 Oct 30;252(1-3):9-16
references.pmid: 18721845
references.citation: Lab Invest. 2008 Oct;88(10):1068-78
references.pmid: 18679376
references.citation: Cancer Res. 2001 Apr 15;61(8):3299-307
references.pmid: 11309284
references.citation: J Biol Chem. 2003 Apr 4;278(14):12029-38
references.pmid: 12556532
references.citation: Proc Natl Acad Sci U S A. 1990 May;87(10):3826-30
references.pmid: 2160079
references.citation: Eur J Biochem. 1992 Mar 1;204(2):491-9
references.pmid: 1371749
references.citation: Carcinogenesis. 2003 Apr;24(4):727-37
references.pmid: 12727802
references.citation: Science. 2009 May 22;324(5930):1076-80
references.pmid: 19461003
references.citation: Science. 1983 Sep 23;221(4617):1256-64
references.pmid: 6351251
references.citation: J Biol Chem. 1990 Aug 25;265(24):14648-53
references.pmid: 2387873
references.citation: Nat Rev Mol Cell Biol. 2006 May;7(5):373-8
references.pmid: 16550215
references.citation: J Biol Chem. 1997 Mar 14;272(11):7445-54
references.pmid: 9054446
references.citation: Hepatology. 2004 May;39(5):1267-76
references.pmid: 15122755
references.citation: Ann Intern Med. 2003 Apr 15;138(8):644-50
references.pmid: 12693887
references.citation: J Biol Chem. 1991 Jun 25;266(18):11632-9
references.pmid: 1646813
references.citation: Biochem J. 2001 Jul 1;357(Pt 1):241-7
references.pmid: 11415455
references.citation: Gastroenterology. 2008 Apr;134(4):1159-68
references.pmid: 18395094
references.citation: Toxicol Sci. 2006 Mar;90(1):111-9
references.pmid: 16352618
references.citation: J Biol Chem. 1999 Nov 19;274(47):33627-36
references.pmid: 10559251
references.citation: Proc Natl Acad Sci U S A. 1997 May 13;94(10):5361-6
references.pmid: 9144242
references.citation: Toxicol Sci. 2001 Jan;59(1):169-77
references.pmid: 11134556
references.citation: Proc Nutr Soc. 2010 Feb;69(1):91-4
references.pmid: 19968906
references.citation: Phytomedicine. 2009 May;16(5):451-5
references.pmid: 19131229
references.citation: J Biol Chem. 2003 Mar 7;278(10):8135-45
references.pmid: 12506115
references.citation: Toxicol Appl Pharmacol. 1995 Nov;135(1):45-57
references.pmid: 7482539
references.citation: Biochem Soc Trans. 2000 Feb;28(2):33-41
references.pmid: 10816095
references.citation: Eur J Pharmacol. 2009 Oct 12;620(1-3):138-44
references.pmid: 19698707
references.citation: Gene. 2002 Jul 10;294(1-2):1-12
references.pmid: 12234662
references.citation: Crit Rev Biochem Mol Biol. 1995;30(6):445-600
references.pmid: 8770536
references.citation: Electrophoresis. 2000 Jun;21(11):2148-61
references.pmid: 10892726
references.citation: Proc Natl Acad Sci U S A. 2001 Apr 10;98(8):4611-6
references.pmid: 11287661
references.citation: Electrophoresis. 2001 May;22(9):1747-63
references.pmid: 11425230
references.citation: Am J Physiol Gastrointest Liver Physiol. 2010 Feb;298(2):G283-94
references.pmid: 19926817