title: Suppression of oxidative phosphorylation in mouse embryonic fibroblast cells deficient in apurinic/apyrimidinic endonuclease.
abstract: The mammalian apurinic/apyrimidinic (AP) endonuclease 1 (APE1) is an essential DNA repair/gene regulatory protein. Decrease of APE1 in cells by inducible shRNA knockdown or by conditional gene knockout caused apoptosis. Here we succeeded in establishing a unique mouse embryonic fibroblast (MEF) line expressing APE1 at a level far lower than those achieved with shRNA knockdown. The cells, named MEF(la) (MEF(lowAPE1)), were hypersensitive to methyl methanesulfonate (MMS), and showed little activity for repairing AP-sites and MMS induced DNA damage. While these results were consistent with the essential role of APE1 in repair of AP sites, the MEF(la) cells grew normally and the basal activation of poly(ADP-ribose) polymerases in MEF(la) was lower than that in the wild-type MEF (MEF(wt)), indicating the low DNA damage stress in MEF(la) under the normal growth condition. Oxidative phosphorylation activity in MEF(la) was lower than in MEF(wt), while the glycolysis rates in MEF(la) were higher than in MEF(wt). In addition, we observed decreased intracellular oxidative stress in MEF(la). These results suggest that cells with low APE1 reversibly suppress mitochondrial respiration and thereby reduce DNA damage stress and increases the cell viability.
authors: Graduate Center for Toxicology, University of Kentucky, Lexington, KY 40536 USA.
authors: Rangaswamy
authors: R
authors: Suganya
authors: Department of Internal Medicine, University of Texas Medical Branch, TX 77555, USA.
authors: Anirban
authors: A
authors: Chakraborty
authors: Graduate Center for Toxicology, University of Kentucky, Lexington, KY 40536 USA; Department of Cellular Biology and Anatomy, LSU Health Sciences Center, Shreveport, LA 71130 USA.
authors: Sumitra
authors: S
authors: Miriyala
authors: Department of Internal Medicine, University of Texas Medical Branch, TX 77555, USA.
authors: Tapas K
authors: TK
authors: Hazra
authors: Graduate Center for Toxicology, University of Kentucky, Lexington, KY 40536 USA. Electronic address: t.izumi@uky.edu.
authors: Tadahide
authors: T
authors: Izumi
mesh_terms: D000818:Animals; D002460:Cell Line / Q000378:metabolism*; D019069:Cell Respiration; D002470:Cell Survival; D002478:Cells, Cultured; D004260:DNA Repair; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism*; D005347:Fibroblasts; D005786:Gene Expression Regulation; D006019:Glycolysis; D045325:HCT116 Cells; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D008928:Mitochondria / Q000378:metabolism; D018384:Oxidative Stress; D010766:Phosphorylation; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism
chemical_list: D011065:Poly(ADP-ribose) Polymerases; C085108:APEX1 protein, human; C486216:Apex1 protein, mouse; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase
keywords: AP endonuclease 1; DNA repair; Endogenous DNA damage; Oxidative phosphorylation
references: Nucleic Acids Res. 1994 Nov 25;22(23):4884-9
references: 7800476
references: Nucleic Acids Res. 1996 Apr 15;24(8):1465-71
references: 8628679
references: Proc Natl Acad Sci U S A. 1996 Aug 20;93(17):8919-23
references: 8799128
references: Proc Natl Acad Sci U S A. 1998 Apr 28;95(9):5061-6
references: 9560228
references: Nat Rev Drug Discov. 2010 Jun;9(6):447-64
references: 20467424
references: Methods. 2010 Aug;51(4):452-7
references: 20347038
references: Cell Mol Life Sci. 2010 Nov;67(21):3589-608
references: 20706766
references: Cell Mol Life Sci. 2010 Nov;67(21):3633-47
references: 20844920
references: Nature. 2011 Feb 17;470(7334):419-23
references: 21278727
references: Pediatr Cardiol. 2011 Mar;32(3):323-8
references: 21286700
references: J Biol Chem. 2011 Sep 16;286(37):31975-83
references: 21768646
references: Nucleic Acids Res. 2011 Oct;39(18):8017-28
references: 21727086
references: Mol Biol Cell. 2011 Oct;22(20):3887-901
references: 21865600
references: J Biol Chem. 2012 Jan 20;287(4):2819-29
references: 22130663
references: Adv Exp Med Biol. 2012;942:269-86
references: 22399427
references: Prog Mol Biol Transl Sci. 2012;107:355-415
references: 22482456
references: J Hypertens. 2012 May;30(5):917-25
references: 22441348
references: J Pharmacol Exp Ther. 2012 Jun;341(3):775-83
references: 22431205
references: DNA Repair (Amst). 2012 Jun 1;11(6):570-8
references: 22497777
references: PLoS Genet. 2012;8(6):e1002689
references: 22685414
references: Free Radic Biol Med. 2012 Jul 15;53(2):237-48
references: 22580151
references: Free Radic Biol Med. 2012 Oct 1;53(7):1478-88
references: 22709585
references: J Neurol Sci. 2012 Nov 15;322(1-2):254-62
references: 22669122
references: Cancer Cell. 2013 Mar 18;23(3):316-31
references: 23453623
references: Cell. 2013 May 9;153(4):910-8
references: 23643243
references: Cell. 2013 Sep 12;154(6):1370-9
references: 23992847
references: Cell Res. 2013 Oct;23(10):1163-71
references: 23979020
references: Nat Protoc. 2013 Nov;8(11):2281-308
references: 24157548
references: Development. 2014 Jan;141(2):269-80
references: 24335252
references: Antioxid Redox Signal. 2014 Feb 1;20(4):708-26
references: 23901781
references: Cell. 2014 May 8;157(4):882-96
references: 24813611
references: Cell Rep. 2014 Sep 25;8(6):1819-31
references: 25220464
references: Nature. 2000 Jan 27;403(6768):451-6
references: 10667800
references: Mutat Res. 2000 Oct 16;461(2):83-108
references: 11018583
references: Cancer Res. 2001 Jul 15;61(14):5552-7
references: 11454706
references: Science. 2002 Feb 22;295(5559):1526-8
references: 11859195
references: Proc Natl Acad Sci U S A. 2002 Mar 19;99(6):3523-8
references: 11904416
references: EMBO J. 2002 Jun 3;21(11):2833-41
references: 12032096
references: J Biol Chem. 2002 Aug 23;277(34):30417-20
references: 12097317
references: Toxicology. 2003 Nov 15;193(1-2):43-65
references: 14599767
references: Nat Rev Cancer. 2004 Jul;4(7):551-61
references: 15229480
references: Mol Cell. 2004 Jul 23;15(2):209-20
references: 15260972
references: Biochemistry. 2004 Sep 14;43(36):11596-604
references: 15350146
references: Basic Life Sci. 1975;5A:31-8
references: 242315
references: J Biol Chem. 1981 Apr 10;256(7):3405-14
references: 6259165
references: J Bacteriol. 1986 Dec;168(3):1120-7
references: 2430946
references: Gene. 1988 May 15;65(1):135-9
references: 3165082
references: J Bacteriol. 1989 May;171(5):2542-6
references: 2468648
references: Nucleic Acids Res. 1989 Aug 11;17(15):6419
references: 2771659
references: Biochemistry. 1992 Apr 14;31(14):3703-8
references: 1567824
references: J Bacteriol. 1992 Dec;174(23):7711-6
references: 1280256
references: Nature. 1993 Apr 22;362(6422):709-15
references: 8469282
references: Bioessays. 1998 Jul;20(7):532-5
references: 9723001
references: Genes Dev. 1998 Oct 1;12(19):3137-43
references: 9765213
references: Mutat Res. 1998 Oct 21;409(1):17-29
references: 9806499
references: J Mol Biol. 1999 Mar 19;287(1):47-57
references: 10074406
references: Biochem J. 1999 Sep 1;342 ( Pt 2):249-68
references: 10455009
references: J Biol Chem. 2008 Sep 26;283(39):26349-56
references: 18635552
references: Proteomics. 2009 Feb;9(4):1058-74
references: 19180539
references: Oncogene. 2009 Apr 2;28(13):1616-25
references: 19219073
references: Antioxid Redox Signal. 2009 Mar;11(3):621-38
references: 18715144
references: Antioxid Redox Signal. 2009 Mar;11(3):601-20
references: 18976116
references: Cancer Res. 2010 Apr 1;70(7):2799-808
references: 20332233
references: Mol Cell. 2005 Feb 4;17(3):463-70
references: 15694346
references: Proc Natl Acad Sci U S A. 2005 Apr 19;102(16):5739-43
references: 15824325
references: J Mol Med (Berl). 2006 Jan;84(1):57-64
references: 16328206
references: Methods Mol Biol. 2006;314:183-99
references: 16673882
references: Head Neck. 2007 Feb;29(2):163-88
references: 17312569
references: Mutat Res. 1994 Jul;315(1):55-63
references: 7517011