© 1995 Oxford University Press
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Synthesis and characterization of covalent adducts derived from the binding of benzo[a]pyrene diol epoxide to a -GGG- sequence in a deoxyoligonucleotide
Chemistry Department and The Radiation and Solid State Laboratory, New York University New York, NY 10003, USA
1To whom correspondence should be addressed
Direct synthesis and purification procedures are described for the preparation of adducts derived from the covalent binding of 7R,8S-dihydroxy-9S,10R-epoxy-7,8,9,10-tetrahydro-benzo[a]pyrene [(+)-ante-BPDE or (+)-BPDE 2] to each of the three guanine residues (trans-N2 lesions) in the oligodeoxyribonucleotide d(CTATG1G2G3TATC) The positions of the modified Gs are defined by Maxam-Gilbert sequencing techniques. Six different oligonucleotides with one or two precisely positioned (+)-anti-BPDE residues are identified. The absorbance, circular dichroism and fluorescence characteristics are changed upon formation of duplexes with the complementary strands d(GATACCCATAG). In the doubly-modified oligonucleotides, a broad, excimer-like long wavelength fluorescence emission band is observed with a maximum near 455 nm only if the two (+)-anti-BPDE-modified Gs are adjacent to one another. The covalently attached (+)-anti-BPDE residues decrease the thermodynamic stabilities of the duplexes; their melting points are markedly dependent on the position of the lesions, being highest with the (+)-anti-BPDE residue at G1 (Tm=40°C, only 2°C lower than in the case of the unmodified oligonucleotide) and lowest when it is situated at G3 (Tm=29°C). The implications of these and other physical characteristics are discussed. The facile synthesis of these or similar site-specific and stereochemically defined (+)-trans-anti-BPDE-N2-dG lesions in runs of contiguous guanines in oligodeoxyribonucleotides of specified base sequence should be useful for the design of site-directed mutagenesis studies in vitro and in vivo.
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