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2006 | 53 | 2 | 337-347
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Sequence-specific p53 gene damage by chloroacetaldehyde and its repair kinetics in Escherichia coli

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Oxidative stress and certain environmental carcinogens, e.g. vinyl chloride and its metabolite chloroacetaldehyde (CAA), introduce promutagenic exocyclic adducts into DNA, among them 1,N6-ethenoadenine (εA), 3,N4-ethenocytosine (εC) and N2,3-ethenoguanine (εG). We studied sequence-specific interaction of the vinyl-chloride metabolite CAA with human p53 gene exons 5-8, using DNA Polymerase Fingerprint Analysis (DPFA), and identified sites of the highest sensitivity. CAA-induced DNA damage was more extensive in p53 regions which revealed secondary structure perturbations, and were localized in regions of mutation hot-spots. These perturbations inhibited DNA synthesis on undamaged template. We also studied the repair kinetics of CAA-induced DNA lesions in E. coli at nucleotide resolution level. A plasmid bearing full length cDNA of human p53 gene was modified in vitro with 360 mM CAA and transformed into E. coli DH5α strain, in which the adaptive response system had been induced by MMS treatment before the cells were made competent. Following transformation, plasmids were re-isolated from transformed cultures 35, 40, 50 min and 1-24 h after transformation, and further subjected to LM-PCR, using ANPG, MUG and Fpg glycosylases to identify the sites of DNA damage. In adaptive response-induced E. coli cells the majority of DNA lesions recognized by ANPG glycosylase were removed from plasmid DNA within 35 min, while MUG glycosylase excised base modifications only within 50 min, both in a sequence-dependent manner. In non-adapted cells resolution of plasmid topological forms was perturbed, suggesting inhibition of one or more bacterial topoisomerases by unrepaired ε-adducts. We also observed delayed consequences of DNA modification with CAA, manifesting as secondary DNA breaks, which appeared 3 h after transformation of damaged DNA into E. coli, and were repaired after 24 h.

Physical description
  • Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland
  • Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland
  • Groupe "Reparation de l'ADN", Institut Gustave Roussy, France
  • Groupe "Reparation de l'ADN", Institut Gustave Roussy, France
  • Institute of Genetics and Biotechnology, Warsaw University, Warszawa, Poland
  • Barbin A, Laib RJ, Bartsch H (1985) Lack of miscoding properties of 7-(2-oxoethyl)guanine, the major vinyl chloride-DNA adduct. Cancer Res 45: 2440-2444.
  • Barbin A, Froment O, Boivin S, Marion MJ, Belpoggi F, Maltoni C, Montesano R (1997) p53 gene mutation pattern in rat liver tumors induced by vinyl chloride. Cancer Res 57: 1695-1698.
  • Bartsch H, Barbin A, Marion MJ, Nair J, Guichard Y (1994) Formation, detection, and role in carcinogenesis of ethenobases in DNA. Drug Metab Rev 26: 349-371.
  • Basu AK, Wood ML, Niedernhofer LJ, Ramos LA, Essigmann JM (1993) Mutagenic and genotoxic effects of three vinyl chloride-induced DNA lesions: 1,N6-ethenoadenine, 3,N4-ethenocytosine, and 4-amino-5-(imidazol-2-yl)imidazole. Biochemistry 32: 12793-12801.
  • Boiteux S, O'Connor TR, Laval J (1987) Formamidopyrimidine-DNA glycosylase of Escherichia coli: cloning and sequencing of the fpg structural gene and overproduction of the protein. EMBO J 6: 3177-3183.
  • Borys-Brzywczy E, Arczewska KD, Saparbaev M, Hardeland U, Schär P, Kuśmierek JT (2005) Mismatch dependent uracil/thymine-DNA glycosylases excise exocyclic hydroxyethano and hydroxypropano cytosine adducts. Acta Biochim Polon 52: 149-165.
  • Bouziane M, Miao F, Ye N, Holmquist G, Chyzak G, O'Connor TR (1998) Repair of DNA alkylation damage. Acta Biochim Polon 45: 191-202.
  • Chen JM, Smith SJ, Marion MJ, Pincus MR, Brandt-Rauf PW (1999) Common conformational effects in the p53 protein of vinyl chloride-induced mutations. J Protein Chem 18: 467-472.
  • Cheng KC, Preston BD, Cahill DS, Dosanjh MK, Singer B, Loeb LA (1991) The vinyl chloride DNA derivative N2,3-ethenoguanine produces G to A transitions in Escherichia coli. Proc Natl Acad Sci USA 88: 9974-9978.
  • Chung FL, Chen HJ, Nath RG (1996) Lipid peroxidation as a potential endogenous source for the formation of exocyclic DNA adducts. Carcinogenesis 17: 2105-2111.
  • Coates PJ, Lorimore SA, Wright EG (2004) Damaging and protective cell signaling in the untargeted effects of ionizing radiation. Mutat Res 568: 5-20.
  • Creech J, Johnson LMN (1974) Angiosarcoma of liver in the manufacture of polyvinyl chloride. J Occup Med 16: 150-151.
  • Delaney JC, Smeester L, Wong C, Frick LE, Taghizadeh K, Wishnok JS, Drennan CL, Samson LD, Essigmann JM (2005) AlkB reverses etheno DNA lesions caused by lipid oxidation in vitro and in vivo. Nat Struct Mol Biol 12: 855-860.
  • Green T, Hathway DE (1978) Interactions of vinyl chloride with rat-liver DNA in vivo. Chem Biol Interact 22: 211-224.
  • Grzesiuk E, Gozdek A, Tudek B (2001) Contribution of E. coli AlkA, TagA glycosylases and UvrABC-excinuclease in MMS mutagenesis. Mutat Res 480-481: 77-84.
  • Guengerich FP, Persmark M, Humphreys WG (1993) Formation of 1,N2- and N3,3-ethenoguanine from 2-halooxiranes: isotopic labeling studies and isolation of hemiaminal derivative of N2-(2-oxoethyl)guanine. Chem Res Toxicol 6: 635-648.
  • Hang B, Chenna A, Guliaev AB, Singer B (2003) Miscoding properties of 1,N6-ethanoadenine, a DNA adduct derived from reaction with the antitumor agent 1,3-bis(2-chloroethyl)-1-nitrosourea. Mutat Res 531: 191-203.
  • Hollstein M, Rice K, Greenblatt MS, Soussi T, Fuchs R, Sorlie T, Hovig E, Smith-Sorensen B, Montesano R, Harris CC (1994a) Database of p53 gene somatic mutations in human tumors and cell lines. Nucleic Acids Res 22: 3551-3555.
  • Hollstein M, Marion MJ, Lehman T, Welsh J, Harris CC, Martel-Planche G, Kusters I, Montesano R (1994b) p53 mutations at A:T base pairs in angiosarcomas of vinyl chloride-exposed factory workers. Carcinogenesis 15: 1-3.
  • Jurado J, Maciejewska A, Krwawicz J, Laval J, Saparbaev M (2004) Role of mismatch-specific uracil-DNA glycosylase in repair of 3,N4-ethenocytosine in vivo. DNA Repair (Amst). 3: 1579-1590.
  • Kadhim MA, Moore SR, Goodwin EH (2004) Interrelationship amongst radiation-induced genomic instability, bystander effects, and the adaptive response. Mutat Res 568: 21-32.
  • Kuśmierek JT, Singer B (1992) 1,N2-ethenodeoxyguanosine: properties and formation in chloroacetaldehyde-treated polynucleotides and DNA. Chem Res Toxicol 5: 634-638.
  • Kuśmierek JT, Folkman W, Singer B (1989) Synthesis of N2,3-ethenodeoxyguanosine, N2,3-ethenodeoxyguanosine 5'-phosphate, and N2,3-ethenodeoxyguanosine 5'-triphosphate. Stability of the glycosyl bond in the monomer and in poly (dG, epsilon dG-dC). Chem Res Toxicol 2: 230-233.
  • Langouët S, Muller M, Guengerich FP (1997) Misincorporation of dNTPs opposite 1,N2-ethenoguanine and 5,6,7,9-tetrahydro-7-hydroxy-9-oxoimidazo[1,2-a]purine in oligonucleotides by Escherichia coli polymerases I exo-and II exo-, T7 polymerase exo- human immunodeficiency virus-1 reverse transcriptase, and rat polymerase beta. Biochemistry 36: 6069-6079.
  • Langouët S, Mican AN, Muller M, Fink SP, Marnett LJ, Muhle SA, Guengerich FP (1998) Misincorporation of nucleotides opposite five-membered exocyclic ring guanine derivatives by Escherichia coli polymerases in vitro and in vivo: 1,N2-ethenoguanine, 5,6,7,9-tetrahydro-9-oxoimidazo[1,2-a]purine, and 5,6,7,9-tetrahydro-7-hydroxy-9-oxoimidazo[1,2-a]purine. Biochemistry 37: 5184-5193.
  • Lee DH, Jin SG, Cai S, Chen Y, Pfeifer GP, O'Connor TR (2005) Repair of methylation damage in DNA and RNA by mammalian AlkB homologues. J Biol Chem. 280: 39448-39459.
  • Levine RL, Yang IY, Hossain M, Pandya GA, Grollman AP, Moriya M (2000) Mutagenesis induced by a single 1,N6-ethenodeoxyadenosine adduct in human cells. Cancer Res 60: 4098-4104.
  • Lilley DMJ (1986) Cyclic adduct formation at structural perturbations in supercoiled DNA molecules. In The Role of Cyclic Nucleic Acids Adducts in Carcinogenesis and Mutagenesis (Singer B, Bartsch, H, eds) pp 83-99, IARC Sci Publ (70) Lyon, IARC.
  • Litinski V, Chenna A, Sagi J, Singer B (1997) Sequence context is an important determinant in the mutagenic potential of 1,N6-ethenodeoxyadenosine (εA): formation of εA base pairs and elongation in defined templates. Carcinogenesis 18: 1609-1615.
  • Maltoni C, Lefemine G, Chieco P, Carretti D (1974) Vinyl chloride carcinogenesis: current results and perspectives. Med Lav 65: 421-444.
  • Matijasevic Z, Sekiguchi M, Ludlum DB (1992) Release of N2,3-ethenoguanine from chloroacetaldehyde-treated DNA by Escherichia coli 3-methyladenine DNA glycosylase II. Proc Natl Acad Sci USA 89: 9331-9334.
  • Mishina Y, Yang CG, He C (2005) Direct repair of the exocyclic DNA adduct 1,N6-ethenoadenine by the DNA repair AlkB proteins. J Am Chem Soc 127: 14594-14595.
  • Moriya M, Zhang W, Johnson F, Grollman AP (1994) Mutagenic potency of exocyclic DNA adducts: marked differences between Escherichia coli and simian kidney cells. Proc Natl Acad Sci USA 91: 11899-11903.
  • Muller M, Belas FJ, Blair IA, Guengerich FP (1997) Analysis of 1,N2-ethenoguanine and 5,6,7,9-tetrahydro-7-hydroxy-9-oxoimidazol[1,2-a]purine in DNA treated with 2-chlorooxirane by high performance liquid chromatography/electrospray mass spectrometry and comparison of amounts to other DNA adducts. Chem Res Toxicol 10: 242-247.
  • O'Connor TR, Boiteux S, Laval J (1988) Ring-opened 7-methylguanine residues in DNA are a block to in vitro DNA synthesis. Nucleic Acid Res 16: 5879-5894.
  • Palejwala VA, Simha D, Humayun MZ (1991) Mechanisms of mutagenesis by exocyclic DNA adducts. Transfection of M13 viral DNA bearing a site-specific adduct shows that ethenocytosine is a highly efficient. Biochemistry 30: 8736-8743.
  • Pandya GA, Moriya M (1996) 1,N6-ethenodeoxyadenosine, a DNA adduct highly mutagenic in mammalian cells. Biochemistry 35: 11487-11492.
  • Pfeifer GP, Dammann R (1999) Measuring the formation and repair of UV photoproducts by ligation-mediated PCR. Methods Mol Biol 113: 213-226.
  • Pourquier P, Bjornsti MA, Pommier Y (1998) Induction of topoisomerase I cleavage complexes by the vinyl chloride adduct 1,N6-ethenoadenine. J Biol Chem 273: 27245-27249.
  • Premaratne S, Mandel M, Mower HF (1993) Identification of DNA adducts at specific locations by sequencing techniques. Int J Biochem 25: 1669-1672.
  • Puisieux A, Lim S, Groopman J, Ozturk M (1991) Selective targeting of p53 gene mutational hotspots in human cancers by etiologically defined carcinogens. Cancer Res 51: 6185-6189.
  • Sabourin M, Osheroff N (2000) Sensitivity of human type II topoisomerases to DNA damage: stimulation of enzyme-mediated DNA cleavage by abasic, oxidized and alkylated lesions. Nucleic Acids Res 28: 1947-1954.
  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press.
  • Saparbaev M, Laval J (1994) Excision of hypoxanthine from DNA containing dIMP residues by the Escherichia coli, yeast, rat, and human alkylpurine DNA glycosylases. Proc Natl Acad Sci USA 91: 5873-5877.
  • Saparbaev M, Kleibl K, Laval J (1995) Escherichia coli, Saccharomyces cerevisiae, rat and human 3-methyladenine DNA glycosylases repair 1,N6-ethenoadenine when present in DNA. Nucleic Acids Res 23: 3750-3755.
  • Saparbaev M, Laval J (1998) 3,N4-ethenodeoxycytidine, a highly mutagenic adduct, is a primary substrate for Escherichia coli double-stranded uracil-DNA-glycosylase. Proc Natl Acad Sci USA 95: 8508-8513.
  • Saparbaev M, Langouët S, Privezentzev CV, Guengerich FP, Cai H, Elder RH, Laval J (2002) 1,N2-ethenoguanine, a mutagenic DNA adduct, is a primary substrate of Escherichia coli mismatch-specific uracil-DNA glycosylase and human alkylpurine-DNA-N-glycosylase. J Biol Chem 277: 26987-26993.
  • Singer B, Kuśmierek JT, Folkman W, Chavez F, Dosanjh MK (1991) Evidence for the mutagenic potential of the vinyl chloride induced adduct, N2,3-ethenodeoxyguanosine, using a site-directed kinetic assay. Carcinogenesis 12: 745-747.
  • Speina E, Cieśla JM, Wójcik J, Bajek M, Kuśmierek JT, Tudek B (2001) The pyrimidine ring-opened derivative of 1,N6-ethenoadenine is excised from DNA by the Escherichia coli Fpg and Nth proteins. J Biol Chem 276: 21821-21827.
  • Speina E, Kierzek AM, Tudek B (2003) Chemical rearrangement and repair pathways of 1,N6-ethenoadenine. Mutat Res 531: 205-217.
  • Tornaletti S, Pfeifer GP (1994) Slow repair of pyrimidine dimers at p53 mutation hotspots in skin cancer. Science 263: 1436-1438.
  • Tudek B, Van Zeeland AA, Kuśmierek JT, Laval J (1998) Activity of Escherichia coli DNA-glycosylases on DNA damaged by methylating and ethylating agents and influence of 3-substituted adenine derivatives. Mutat Res 407: 169-176.
  • Tudek B, Kowalczyk P, Cieśla JM (1999) Localisation of chloroacetaldehyde-induced DNA damage in human p53 gene by DNA polymerase fingerprint analysis. In Exocyclic DNA Adducts in Mutagenesis and Carcinogenesis (Singer B, Bartsch H, eds) pp 279-293, No. 150. IARC Scientific Publications, Lyon IARC.
  • Velez-Cruz R, Riggins JN, Daniels JS, Cai H, Guengerich FP, Marnett LJ, Osherhoff N (2005) Exocyclic DNA lesions stimulate DNA cleavage mediated by human topoisomerase IIα in vitro and in cultured cells. Biochemistry 44: 3972-3981.
  • Viola PL (2001) Pathology of vinyl chloride. Med Lav 92: 509-515.
  • Volkert MR (1988) Adaptive response of Escherichia coli to alkylation damage. Environ Mol Mutagen 11: 241-255.
  • Xia L, Zheng L, Lee HW, Bates SE, Federico L, Shen B, O'Connor TR (2005) Human 3-methyladenine-DNA glycosylase; effect of sequence context on excision, association with PCNA, and stimulation by AP endonuclease. J Mol Biol 346: 1259-1274.
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