title: Sequence- and stereospecific conformational rearrangement of styrene oxide adducts located at A x C mismatched base pairs.
abstract: The solution structures of R- and S-alpha-(N(6)-adenyl)-styrene oxide adducts mismatched with cytosine at position X(7) in d(CGGACAXGAAG) x d(CTTCCTGTCCG), incorporating codons 60, 61 (underlined), and 62 of the human N-ras protooncogene, were determined. These were the R- and S(61,3)C adducts. The structures for these mismatched adducts differed from the sequence isomeric R- and S(61,2)C adducts [Painter, S. L., Zegar, I. S., Tamura, P. J., Bluhm, S., Harris, C. M., Harris, T. M., and Stone, M. P. (1999) Biochemistry 38, 8635-8646]. The results reveal that the structural consequences of cytosine mispairing opposite the R- and S-alpha-SO adducts differ as a function of DNA sequence. The thermodynamic stability of both the R- and S(61,3)C mismatched adducts was dependent upon pH. At neutral pH, the R- and S(61,3)C adducts exhibited significant structural perturbation and had lower T(m) values, as compared to the R- and S(61,2)C adducts. In both instances, this was attributed to reorientation about the C6-N(6) bond, such that the N(6)H proton faced away from the Watson-Crick face of the purine base and into the major groove. The conformation about the N(6)-C(alpha)-C(beta)-O torsion angle was predicted from rMD calculations to be stabilized by a N/O gauche-type interaction between the styrenyl hydroxyl moiety and adenine N(6) at the lesion site. For the R(61,3)C adduct, the styrenyl moiety remained oriented in the major groove and faced in the 3'-direction. In the properly base-paired R(61,3) adduct, it had faced in the 5' direction. For the S(61,3)C adduct, the styrene ring was inserted into the duplex, approximately perpendicular to the helical axis of the DNA. It faced in the 5'-direction. In the properly base-paired S(61,3) adduct, it had faced in the 3'-direction. The results were correlated with site-specific mutagenesis experiments in vivo. The latter revealed that the R- and S(61,3)-alpha-styrene oxide adducts were nonmutagenic. This may be a consequence of the greater structural perturbation associated with formation of the cytosine mismatch at neutral pH for the R- and S(61,3) adducts as compared to the S(61,2) adduct that exhibited low levels of A --> G mutations.
authors.affiliation: Institute of Organic Chemistry with Center of Phytochemistry, Bulgarian Academy of Sciences, BG 1113 Sofia, Bulgaria. mario@orgchm.bas.bg
authors.forename: M F
authors.initials: MF
authors.lastname: Simeonov
authors.forename: P J
authors.initials: PJ
authors.lastname: Tamura
authors.forename: A S
authors.initials: AS
authors.lastname: Wilkinson
authors.forename: C M
authors.initials: CM
authors.lastname: Harris
authors.forename: T M
authors.initials: TM
authors.lastname: Harris
authors.forename: M P
authors.initials: MP
authors.lastname: Stone
mesh_terms: D000225:Adenine / Q000737:chemistry*; D020137:Base Pair Mismatch; D001483:Base Sequence; D003198:Computer Simulation; D003596:Cytosine / Q000737:chemistry*; D018736:DNA Adducts / Q000737:chemistry*; D004852:Epoxy Compounds / Q000737:chemistry*; D006860:Hydrogen Bonding; D006863:Hydrogen-Ion Concentration; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D009690:Nucleic Acid Conformation; D009692:Nucleic Acid Heteroduplexes / Q000737:chemistry; D011522:Protons; D013237:Stereoisomerism; D013816:Thermodynamics
chemical_list: D018736:DNA Adducts; D004852:Epoxy Compounds; D009692:Nucleic Acid Heteroduplexes; D011522:Protons; D003596:Cytosine; C013690:styrene oxide; D000225:Adenine