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2–Aminofluorene modified DNA duplex exists in two interchangeable conformations

Abstract

One– and two–dimensional NMR shows that the carcinogen 2–aminofluorene exists in two unique, interchangeable conformations when covalently bound to a model human c–H–ras1 proto–oncogene codon 61 oligomer duplex. In one conformation the 2–aminofluorene moiety protrudes out of the major groove leaving the Watson–Crick base pairing of the cytosine and 2–aminofluorene–guanine bases intact, consistent with the ability of replicating enzymes to bypass the lesion and correctly incorporate cytosine. The second form of the modified oligomer duplex may be representative of a pre–mutagenic conformation in that the 2–aminofluorene moiety is stacked within the DNA helix, disrupting base pairing between the 2–aminofluorene–modified guanine and its complementary cytosine.

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References

  1. Kriek, E. Fifty years of research on N-acetyl-2-aminofluorene, one of the most versatile compounds in experimental cancer research. J. Cancer Res. clin. Oncol. 118, 481–489 (1992).

    Article  CAS  Google Scholar 

  2. Miller, J.A. Carcinogenesis by chemicals: An overview — G. H. A. Clowes Memorial Lecture. Cancer Res. 30, 559–576 (1970).

    CAS  PubMed  Google Scholar 

  3. Beland, F.A. & Kadlubar, F.F. in Handbook of Experimental Pharmacology 94/1, 281–285 (eds Cooper, C.S. & Grover, P. L.) (Springer-Verlag, Heidelberg, 1990).

    Google Scholar 

  4. Kriek, E., Miller, J.A., Juhl, U. & Miller, E.C. 8-(N-2-Flurenoacetamido)guanosine, an arylamidation reaction product of guanosine and the carcinogen N-acetoxy-N-2-fluorenylacetamide in neutral solution. Biochemistry 6, 177–182 (1967).

    Article  CAS  Google Scholar 

  5. Irving, C.C. Enzymatic deacetylation of N-hydroxy-2-acetylaminofluorene by liver microsomes Cancer Research 26, 1390–1396 (1966).

    CAS  PubMed  Google Scholar 

  6. Beland, F.A., Dooley, K.L. & Jackson, C.D. Persistance of DNA adducts in rat liver and kidney after multiple doses of the carcinogen N-hydroxy-2-acetylaminofluorene. Cancer Res. 42, 1348–1354 (1982).

    CAS  PubMed  Google Scholar 

  7. Gupta, R.C. & Dighe, N.R. Formation and removal of DNA adducts in rat liver treated with N-hydroxy derivatives of 2-acetylaminofluorene, 4-acetylaminobiphenyl, and 2-acetylaminophenanthrene. Carcinogenesis 5, 343–349 (1984).

    Article  CAS  Google Scholar 

  8. Poirier, M.C., Fullerton, N.F., Kinouchi, T., Smith, B.A. & Beland, F. A. Comparison between DNA adduct formation and tumorigenesis in livers and bladders of mice chronically fed 2-acetylaminofluorene. Carcinogenesis 12, 895–900 (1991).

    Article  CAS  Google Scholar 

  9. Gupta, P.K., Lee, M. & King, C.M. Comparison of mutagenesis induced in single- and double-stranded M13 viral DNA by treatment with N-hydroxy-2-aminofluorene. Carcinogenesis 9, 1337–1345(1988)

    Article  CAS  Google Scholar 

  10. Carothers, A.D. et al A mutational hot spot induced by N-hydroxy-aminofluorene in dihydrofolate reductase mutants of Chinese hamster ovary cells. Carcinogenesis 14, 2181–2184 (1993).

    Article  CAS  Google Scholar 

  11. Mah, M.C., Maher, V.M., Thomas, H., Reid, T.M., King, C.M. & McCormick, J.J. Mutations induced by aminofluorene–DNA adducts during replication in human cells. Carcinogenesis 10, 2321–2328(1989).

    Article  CAS  Google Scholar 

  12. Bichara, M. & Fuchs, R.P.P. DNA Binding and mutation spectra of the carcinogen N-2-aminofluorene in Escherichia coli: A correlation between the conformation of the premutagenic lesion and the mutagenic specificity. J. molec. Biol. 183, 341–351 (1985).

    Article  CAS  Google Scholar 

  13. Shibutani, S. & Grollman, A.P. On the mechanism of frameshift (deletion) mutagenesis in vitro. J. biol. Chem. 268, 11703–11710 (1993).

    CAS  PubMed  Google Scholar 

  14. Michaels, M.L., Reid, T.M., King, C.M. & Romano, L.J. Accurate in vitro translesion synthesis by Escherichia coli DNA polymerase I (large fragment) on a site-specific, aminofluorene-modified oligonucleotide. Carcinogenesis 12, 1641–1646 (1991).

    Article  CAS  Google Scholar 

  15. Lutgerink, J.T. et al Bypass of the major aminofluorene-DNA adduct during in vivo replication of single- and double-stranded fX174 DNA treated with N-hydroxy-2-aminofluorene. Carcinogenesis 6, 1501–1506(1985).

    Article  CAS  Google Scholar 

  16. Santella, R.M., Kreik, E. & Grunberger, D. Circular dichroism and proton magnetic resonance studies of dApdG modified with 2-aminofluorene and 2-acetyla-aminofluorene. Carcinogenesis 1, 897–902 (1980).

    Article  CAS  Google Scholar 

  17. Leng, M., Ptak, M. & Rio, P. Conformation of acetylaminofluorene and aminofluorene modified guanosine and guanosine derivatives. Biochem. Biophys. Res. Comm. 96, 1095–1102(1980).

    Article  CAS  Google Scholar 

  18. Evans, F.E., Miller, D.W. & Beland, F.A. Sensitivity of the conformation of deoxyguanosine to binding at the C-8 position by N-acetylated and unacetylated 2-aminofluorene. Carcinogenesis 1, 955–959 (1980).

    Article  CAS  Google Scholar 

  19. Kriek, E. and Spelt, C.E. Differential excision from DNA of the C-8 deoxyguanosine reaction products of N-hydroxy-2-aminofluorene and N-acetoxy-N-acetyl-2-aminofluorene by endonuclease S1 from Aspergiilus Oryzae. Cancer Lett. 7, 147–154 (1979).

    Article  CAS  Google Scholar 

  20. Sage, E., Spodheim-Maurizot, M., Rio, P., Leng, M. & Fuchs, R.P.P. Discrimination by antibodies between local defects in DNA induced by 2-aminofluorene derivatives. FEBS Lett. 108, 66–68 (1979).

    Article  CAS  Google Scholar 

  21. Hingerty, B.E. & Broyde, S. Energy minimized structures of carcinogen-DNA adducts: 2-acetylaminofluorene and 2-aminofluorene. J. Biomolec. Struc. Dyn. 4, 365–372 (1986).

    Article  CAS  Google Scholar 

  22. Broyde, S. & Hingerty, B. Conformation of 2-aminofluorene-modified DNA. Biopolymers 22, 2423–2441(1983).

    Article  CAS  Google Scholar 

  23. Lipkowitz, K.B., Chevalier, T., Widdifield, M. & Beland, F.A. Force field conformational analysis of aminofluorene and acetylaminofluorene substituted deoxyguanosine. Chem. Biol. Interact. 40, 57–76(1982).

    Article  CAS  Google Scholar 

  24. Norman, D. et al. NMR and computational characterization of the N-(deoxyguanosin-8-yl)aminofluorene adduct [(AF)G] opposite adenosine in DNA: (AF)G[syn]•A[anti] pair formation and its pH dependence. Biochemistry 28, 7462–7476 (1989).

    Article  CAS  Google Scholar 

  25. van Houte, L.P.A., Westra, J.G., Retel, J. & van Grondelle, R.A. Circular dichroism study on the conformation of d(CGT) modified with N-acetyl-2-aminofluorene or 2-aminofluorene. J. Biomolec. Struct. Dyn. 9, 45–59 (1991).

    Article  CAS  Google Scholar 

  26. Wemmer, D.E. . in Biological Magnetic Resonance. 10 (eds Berliner, L. J., & Reuben, J.) 195–264 (Plenum, New York, 1992).

    Google Scholar 

  27. Wüthrich, K. NMR of Proteins and Nucleic Acids (Wiley, New York, 1986).

    Book  Google Scholar 

  28. Skelnár, V., Brooks, B.R., Zon, G. & Bax, A. Absorption mode two-dimensional NOE spectroscopy of exchangeable protons in oligonucleotides. FEBS Lett. 216, 249–252 (1987).

    Article  Google Scholar 

  29. Grunberger, D. & Carothers, A. Changes in DNA conformation and types of mutation induced in CHO dhfr gene by N-2-acetylaminofluoren and N-2-aminofluorene. Collect. Czech. chem. Commun. 56, 1151–1165 (1991).

    Article  CAS  Google Scholar 

  30. Shibutani, S., Takeshita, M. & Grollman, A.P. Insertion of specific bases during DNA synthesis past the oxidation-damaged 8-oxodG. Nature 348, 431–434 (1991).

    Article  Google Scholar 

  31. Ripley, L.S. Frameshift mutation: Determinants of specificity. A. Rev. Genet. 24, 189–213 (1990).

    Article  CAS  Google Scholar 

  32. Kunkel, T.A. Misalignment-mediated DNA synthesis errors. Biochemistry 29, 8003–8011 (1990).

    Article  CAS  Google Scholar 

  33. Belguise-Valladier, P. & Fuchs, R.P.P. Strong sequence-dependent polymorphism in adduct-induced DNA structure: Analysis of single N-2–acetylaminofluorene residues bound within the Nar I mutation hot spot. Biochemistry 30, 10091–10100 (1991).

    Article  CAS  Google Scholar 

  34. Veaute, X. & Fuchs, R.P.P. Polymorphism in N-2-acetylaminofluorene induced DNA structure as revealed by DNase I footprinting. Nucl. Acids Res. 19, 5603–5606 (1991).

    Article  CAS  Google Scholar 

  35. Garcia, A., Lambert, I.B. & Fuchs, R.P.P. DNA adduct-induced stabilization of slipped frameshift intermediates within repetitive sequences: Implications for mutagenesis. Proc. natn. Acad. Sci. U.S.A. 90 5989–5993 (1993).

    Article  CAS  Google Scholar 

  36. Gupta, P.K., Pandrangi, R.G., Lee, M. & King, C.M. Induction of mutations by N-acetoxy-N-acetyl-2-aminofluorene modified M13 viral DNA. Carcinogenesis 12, 819–824 (1991).

    Article  CAS  Google Scholar 

  37. O'Handley, S.F. et al. Structural characterization of an N-acetyl-2-aminofluorene (AAF) modified DNA oligomer by NMR, energy minimization, and molecular dynamics. Biochemistry 32, 2481–2481 (1993).

    Article  CAS  Google Scholar 

  38. Evans, F.E., Miller, D.W. & Levine, R.A. 1H NMR Study of self-association and restricted internal rotation of the C8-substituted deoxyguanosine 5′-monophosphate adduct of the carcinogen 2-(acetylamino)fluorene. J. Biomolec. Struct. Dyn. 3, 935–948 (1986).

    Article  CAS  Google Scholar 

  39. Cho, B.P., Beland, F.A. & Marques, M.M. NMR stuctural studies of a 15-mer DNA duplex from a ras protooncogene modified with the carcinogen 2-Amniofluorene: conformational heterogeneity. Biochemistry (1994) in press.

    Google Scholar 

Download references

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Eckel, L., Krugh, T. 2–Aminofluorene modified DNA duplex exists in two interchangeable conformations. Nat Struct Mol Biol 1, 89–94 (1994). https://doi.org/10.1038/nsb0294-89

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