title: Translesion synthesis past acrolein-derived DNA adducts by human mitochondrial DNA polymerase γ.
abstract: Acrolein, a mutagenic aldehyde, is produced endogenously by lipid peroxidation and exogenously by combustion of organic materials, including tobacco products. Acrolein reacts with DNA bases forming exocyclic DNA adducts, such as γ-hydroxy-1,N(2)-propano-2'-deoxyguanosine (γ-HOPdG) and γ-hydroxy-1,N(6)-propano-2'-deoxyadenosine (γ-HOPdA). The bulky γ-HOPdG adduct blocks DNA synthesis by replicative polymerases but can be bypassed by translesion synthesis polymerases in the nucleus. Although acrolein-induced adducts are likely to be formed and persist in mitochondrial DNA, animal cell mitochondria lack specialized translesion DNA synthesis polymerases to tolerate these lesions. Thus, it is important to understand how pol γ, the sole mitochondrial DNA polymerase in human cells, acts on acrolein-adducted DNA. To address this question, we investigated the ability of pol γ to bypass the minor groove γ-HOPdG and major groove γ-HOPdA adducts using single nucleotide incorporation and primer extension analyses. The efficiency of pol γ-catalyzed bypass of γ-HOPdG was low, and surprisingly, pol γ preferred to incorporate purine nucleotides opposite the adduct. Pol γ also exhibited ∼2-fold lower rates of excision of the misincorporated purine nucleotides opposite γ-HOPdG compared with the corresponding nucleotides opposite dG. Extension of primers from the termini opposite γ-HOPdG was accomplished only following error-prone purine nucleotide incorporation. However, pol γ preferentially incorporated dT opposite the γ-HOPdA adduct and efficiently extended primers from the correctly paired terminus, indicating that γ-HOPdA is probably nonmutagenic. In summary, our data suggest that acrolein-induced exocyclic DNA lesions can be bypassed by mitochondrial DNA polymerase but, in the case of the minor groove γ-HOPdG adduct, at the cost of unprecedented high mutation rates.
authors.affiliation: Mitochondrial DNA Replication Group, Laboratory of Molecular Genetics, NIEHS, National Institutes of Health, Research Triangle Park, North Carolina 27709.
authors.forename: Rajesh
authors.initials: R
authors.lastname: Kasiviswanathan
authors.affiliation: Center for Research on Occupational and Environmental Toxicology, Oregon Health and Science University, Portland, Oregon 97239.
authors.forename: Irina G
authors.initials: IG
authors.lastname: Minko
authors.affiliation: Center for Research on Occupational and Environmental Toxicology, Oregon Health and Science University, Portland, Oregon 97239; Department of Molecular and Medical Genetics, Oregon Health and Science University, Portland, Oregon 97239.
authors.forename: R Stephen
authors.initials: RS
authors.lastname: Lloyd
authors.affiliation: Mitochondrial DNA Replication Group, Laboratory of Molecular Genetics, NIEHS, National Institutes of Health, Research Triangle Park, North Carolina 27709. Electronic address: copelan1@niehs.nih.gov.
authors.forename: William C
authors.initials: WC
authors.lastname: Copeland
mesh_terms: D000171:Acrolein / Q000494:pharmacology*; D020134:Catalytic Domain; D018736:DNA Adducts; D004249:DNA Damage; D000074002:DNA Polymerase gamma; D004261:DNA Replication; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D003849:Deoxyguanosine / Q000737:chemistry; D006801:Humans; D015227:Lipid Peroxidation; D008928:Mitochondria / Q000201:enzymology*; D008956:Models, Chemical; D016296:Mutagenesis; D009841:Oligonucleotides / Q000737:chemistry; D018384:Oxidative Stress
chemical_list: D018736:DNA Adducts; D009841:Oligonucleotides; D000171:Acrolein; D000074002:DNA Polymerase gamma; D004259:DNA-Directed DNA Polymerase; D003849:Deoxyguanosine
keywords: Acrolein; DNA Damage; DNA Polymerase; DNA Replication; Mitochondrial DNA; Oxidative Stress; Translesion DNA Synthesis
references.citation: J Biol Chem. 2001 Mar 23;276(12):9071-6
references.pmid: 11124950
references.citation: Mutat Res. 1999 Mar 8;424(1-2):71-81
references.pmid: 10064851
references.citation: Chem Res Toxicol. 2000 May;13(5):421-9
references.pmid: 10813660
references.citation: Mol Cell Biol. 2005 Oct;25(19):8748-54
references.pmid: 16166652
references.citation: DNA Repair (Amst). 2011 Aug 15;10(8):826-34
references.pmid: 21622032
references.citation: Nucleic Acids Res. 2010 Jun;38(10):3233-44
references.pmid: 20144948
references.citation: Mitochondrion. 2006 Jun;6(3):136-42
references.pmid: 16725382
references.citation: Cancer Res. 1984 Mar;44(3):990-5
references.pmid: 6318992
references.citation: Nature. 2004 Mar 4;428(6978):97-100
references.pmid: 14999287
references.citation: J Biol Chem. 2003 Feb 21;278(8):5970-6
references.pmid: 12502710
references.citation: Chem Res Toxicol. 2010 Mar 15;23(3):689-95
references.pmid: 20102227
references.citation: J Mol Biol. 2003 May 23;329(1):45-57
references.pmid: 12742017
references.citation: J Biol Chem. 2003 Jan 10;278(2):784-90
references.pmid: 12401796
references.citation: Biochim Biophys Acta. 2012 Sep-Oct;1819(9-10):979-91
references.pmid: 22728831
references.citation: Chem Res Toxicol. 2008 Oct;21(10):1983-90
references.pmid: 18788757
references.citation: J Biol Chem. 2001 Mar 23;276(12):9066-70
references.pmid: 11106660
references.citation: Chem Res Toxicol. 2002 Mar;15(3):367-72
references.pmid: 11896684
references.citation: Nucleic Acids Res. 2009 May;37(9):2830-40
references.pmid: 19282446
references.citation: Neurochem Int. 2005 Feb;46(3):243-52
references.pmid: 15670641
references.citation: Mol Nutr Food Res. 2008 Jan;52(1):7-25
references.pmid: 18203133
references.citation: DNA Repair (Amst). 2007 Dec 1;6(12):1732-9
references.pmid: 17689152
references.citation: J Biol Chem. 2002 May 24;277(21):18257-65
references.pmid: 11889127
references.citation: Cell. 2009 Oct 16;139(2):312-24
references.pmid: 19837034
references.citation: Hum Mol Genet. 2009 Oct 15;18(R2):R169-76
references.pmid: 19808793
references.citation: J Biol Chem. 2005 Jan 14;280(2):1165-78
references.pmid: 15533946
references.citation: J Biol Chem. 2001 Oct 12;276(41):38090-6
references.pmid: 11477093
references.citation: Chem Rev. 2006 Feb;106(2):383-405
references.pmid: 16464011
references.citation: Cancer Res. 1996 Feb 1;56(3):452-6
references.pmid: 8564951
references.citation: Biochemistry. 1998 Jul 21;37(29):10529-39
references.pmid: 9671525
references.citation: Methods. 2010 Aug;51(4):379-84
references.pmid: 20176107
references.citation: Proc Natl Acad Sci U S A. 2005 Apr 5;102(14):4990-5
references.pmid: 15784738
references.citation: J Biol Chem. 1995 Apr 21;270(16):9202-6
references.pmid: 7721837
references.citation: J Biol Chem. 2001 Mar 23;276(12):9077-82
references.pmid: 11054428
references.citation: J Biol Chem. 1999 Dec 31;274(53):38197-203
references.pmid: 10608893
references.citation: Hum Mol Genet. 2007 Nov 15;16(22):2729-39
references.pmid: 17725985
references.citation: Mol Cell Biol. 2004 Jul;24(13):5687-93
references.pmid: 15199127
references.citation: J Biol Chem. 2012 Mar 16;287(12):9222-9
references.pmid: 22194617
references.citation: J Biol Chem. 2006 Nov 24;281(47):36241-8
references.pmid: 17005553
references.citation: Environ Health Perspect. 2001 Mar;109(3):219-24
references.pmid: 11333181
references.citation: J Am Chem Soc. 2002 Aug 14;124(32):9324-5
references.pmid: 12166998
references.citation: IARC Sci Publ. 1999;(150):45-54
references.pmid: 10626207
references.citation: Arch Environ Contam Toxicol. 1999 May;36(4):373-6
references.pmid: 10227855
references.citation: J Biol Chem. 1994 Nov 25;269(47):29984-92
references.pmid: 7961998
references.citation: J Am Chem Soc. 2003 Jan 8;125(1):50-61
references.pmid: 12515506
references.citation: J Biol Chem. 2010 Jan 8;285(2):1490-9
references.pmid: 19858216
references.citation: J Bacteriol. 2011 Aug;193(15):3815-21
references.pmid: 21622737
references.citation: J Biol Chem. 1994 Nov 25;269(47):29993-7
references.pmid: 7961999
references.citation: Mutat Res. 2008 Jan 1;637(1-2):161-72
references.pmid: 17868748
references.citation: Chem Res Toxicol. 2009 May;22(5):759-78
references.pmid: 19397281
references.citation: Genetics. 2006 Apr;172(4):2683-8
references.pmid: 16452144
references.citation: J Biol Chem. 1998 Jun 26;273(26):16058-66
references.pmid: 9632657
references.citation: Nucleic Acids Res. 2004 Jan 16;32(1):397-405
references.pmid: 14729924
references.citation: Proc Natl Acad Sci U S A. 1994 Aug 2;91(16):7491-5
references.pmid: 8052609
references.citation: J Biol Chem. 2000 Apr 28;275(17):13017-25
references.pmid: 10777605
references.citation: Mol Nutr Food Res. 2011 Sep;55(9):1401-10
references.pmid: 21809440
references.citation: Mol Cell Biol. 2004 Aug;24(16):6900-6
references.pmid: 15282292
references.citation: Cell. 2010 Apr 16;141(2):280-9
references.pmid: 20403324