Preferences help
enabled [disable] Abstract
Number of results
2005 | 52 | 4 | 867-874
Article title

Characterization of a novel protein that specifically binds to DNA modified by N-acetoxy-acetylaminofluorene and cis-diamminedichloroplatinum

Title variants
Languages of publication
Proteins recognizing DNA damaged by the chemical carcinogen N-acetoxy-acetylaminofluorene (AAAF) were analyzed in nuclear extracts from rat tissues, using a 36 bp oligonucleotide as a substrate and electrophoretic mobility shift and Southwestern blot assays. One major damage-recognizing protein was detected, whose amount was estimated as at least 105 copies per cell. Levels of this protein were similar in extracts from brain, kidney and liver, but much lower in extracts from testis. The affinity of the detected protein for DNA damaged by AAAF was about 70-fold higher than for undamaged DNA. DNA damaged by cis-diamminedichloroplatinum (cis-DDP), benzo(a)pyrene diolepoxide (BPDE) or UV-radiation also bound this protein with an increased affinity, the former more strongly and the latter two more weakly as compared to AAAF-damaged DNA. The detected AAAF/DDP-damaged-DNA-binding (AAAF/DDP-DDB) protein had a molecular mass of about 25 kDa and was distinct from histone H1 or HMGB proteins, which are known to have a high affinity for cis-DDP-damaged DNA. The level of this damage-recognizing protein was not affected in rats treated with the carcinogen 2-acetylaminofluorene. The activity of an AAAF/DDP-DDB protein could also be detected in extracts from mouse liver cells but not from the Hep2G human hepatocellular carcinoma.
Physical description
  • Department of Experimental and Clinical Radiobiology, Maria Skłodowska-Curie Memorial Cancer Center and Institute of Oncology, Gliwice, Poland
  • Department of Experimental and Clinical Radiobiology, Maria Skłodowska-Curie Memorial Cancer Center and Institute of Oncology, Gliwice, Poland
  • Araujo SJ, Wood RD (1999) Protein complexes in nucleotide excision repair. Mutat Res 435: 23-33.
  • Banchev T, Srebreva L, Zlatanova J (1991) Purification of histone H10 and its subfractions under non-denaturing conditions. Biochim Biophys Acta 1073: 230-232.
  • Birger Y, West KL, Postnikov YV, Lim J-H, Furusawa T, Wagner JP, Laufer CS, Kraemer KH, Bustin M (2003) Chromosomal protein HMGN1 enhances the rate of DNA repair in chromatin. EMBO J 22: 1665-1675.
  • Bohr VA, Phillips DH, Hanawalt PC (1987) Heterogeneous DNA damage and repair in the mammalian genome. Cancer Res 47: 6426-6436.
  • Chu G (1994) Cellular responses to cisplatin. The roles of DNA-binding proteins and DNA repair. J Biol Chem 269: 787-790.
  • de Laat WL, Jaspers NG, Hoeijmakers JH (1999) Molecular mechanism of nucleotide excision repair. Genes Dev 13: 768-785.
  • Gupta RC (1985) Enhanced sensitivity of 32P-postlabeling analysis of aromatic carcinogen:DNA adducts. Cancer Res 45: 5656-5662.
  • Heflich RH, Neft RE (1994) Genetic toxicity of 2-acetylaminofluorene, 2-aminofluorene and some of their metabolites and model metabolites. Mutat Res 318: 73-114.
  • Kroese ED, van de Poll ML, Mulder GJ, Meerman JH (1988) The role of N-sulfation in the N-hydroxy-2-acetylaminofluorene-mediated outgrowth of diethylnitrosamine-initiated hepatocytes to gamma-glutamyltranspeptidase-positive foci in male rat liver. Carcinogenesis 9: 1953-1958.
  • Łanuszewska J, Widłak P (2000) High mobility group 1 and 2 proteins bind preferentially to DNA that contains bulky adducts induced by benzo(a)pyrene diol epoxide and N-acetoxy-acetylaminofluorene. Cancer Lett 158: 17-25.
  • McGregor WG, Wei D, Chen RH, Maher VM, McCormick JJ (1997) Relationship between adduct formation, rates of excision repair and the cytotoxic and mutagenic effects of structurally-related polycyclic aromatic carcinogens. Mutat Res 376: 143-152.
  • Pasheva EA, Pashev IG, Favre A (1998) Preferential binding of high mobility group 1 protein to UV-damaged DNA. Role of the COOH-terminal domain. J Biol Chem 273: 24730-24736.
  • Petit C, Sancar A (1999) Nucleotide excision repair: from E. coli to man. Biochimie 81: 15-25.
  • Pietrowska M, Łanuszewska J, Walter Z, Rzeszowska-Wolny J, Widłak P (2000) Detection and characterization of rat protein recognizing DNA damaged by N-acetoxy-acetylaminofluorene. Cell Mol Biol Lett 5: 423-431.
  • Pil PM, Lippard SJ (1992) Specific binding of chromosomal protein HMG1 to DNA damaged by the anticancer drug cisplatin. Science 256: 234-237.
  • Poirier MC, Fullerton NF, Kinouchi T, Smith BA, Beland FA (1991) Comparison between DNA adduct formation and tumorigenesis in livers and bladders of mice chronically fed 2-acetylaminofluorene. Carcinogenesis 12: 895-900.
  • Protic M, Levine AS (1993) Detection of DNA damage-recognition proteins using the band-shift assay and southwestern hybridization. Electrophoresis 14: 682-692.
  • Protic M (1994) Eukaryotic damaged DNA-binding proteins. DNA repair proteins or transcription factors? Ann N Y Acad Sci 726: 333-335.
  • Rzeszowska-Wolny J, Widlak P (1999) Damaged DNA-binding proteins: recognition of N-acetoxy-acetylaminofluorene-induced DNA adducts. Acta Biochim Polon 46: 173-180.
  • Spiker S (1984) High-mobility group chromosomal proteins of wheat. J Biol Chem 259: 12007-12013.
  • Stein A (1989) Reconstitution of chromatin from purified components. Methods Enzymol 170: 585-603.
  • Tang MS, Bohr VA, Zhang XS, Pierce J, Hanawalt PC (1989) Quantification of aminofluorene adduct formation and repair in defined DNA sequences in mammalian cells using the UVRABC nuclease. J Biol Chem 264: 14455-14462.
  • Ushay HM, Tullius TD, Lippard SJ (1981) Inhibition of the BamHI cleavage and unwinding of pBR322 deoxyribonucleic acid by the antitumor drug cis-dichlorodiammine-platinum(II). Biochemistry 20: 3744-3748.
  • Vaisman A, Chaney SG (1995) Induction of UV-damage recognition protein by cisplatin treatment. Biochemistry 34: 105-114.
  • Wang JF, Bashir M, Engelsberg BN, Witmer C, Rozmiarek H, Billings PC (1997) High mobility group proteins 1 and 2 recognize chromium-damaged DNA. Carcinogenesis 18: 371-375.
  • Widłak P, Rzeszowska-Wolny J (1999) Nuclear matrix and nucleotide excision repair: damage-recognition proteins are not constitutive components of the nuclear matrix. In: Gene Therapy and Molecular Biology; vol. 4, Boulikas T, ed, pp 275-284. Gene Therapy Press, Palo Alto.
  • Wood RD (1999) DNA damage recognition during nucleotide excision repair in mammalian cells. Biochimie 81: 39-44.
  • Yaneva J, Leuba SH, van Holde K, Zlatanova J (1997) The major chromatin protein histone H1 binds preferentially to cis-platinum-damaged DNA. Proc Natl Acad Sci USA 94: 13448-13451.
Document Type
Publication order reference
YADDA identifier
JavaScript is turned off in your web browser. Turn it on to take full advantage of this site, then refresh the page.