Influence of the R(61,2)- and S(61,2)-alpha-(N6-adenyl)styrene oxide adducts on the A.C mismatched base pair in an oligodeoxynucleotide containing the human N-ras codon 61.
Conformational studies of R- and S-alpha-(N6-adenyl)styrene oxide adducts mismatched with deoxycytosine at position X6 in d(CGGACXAGAAG).d(CTTCTCGTCCG), incorporating codons 60, 61 (underlined), and 62 of the human N-ras protooncogene, are described. These were the R- and S(61,2)C adducts. The S(61,2)C adduct afforded a stable solution structure, while the R(61,2)C adduct resulted in a disordered structure. Distance restraints for the S(61, 2)C adduct were calculated from NOE data using relaxation matrix analysis. These were incorporated as effective potentials into the total energy equation. The structures were refined using restrained molecular dynamics calculations which incorporated a simulated annealing protocol. The accuracy of the emergent structures was evaluated by complete relaxation matrix methods. The structures refined to an average rms difference of 1.07 A, determined by pairwise analysis. The experimentally determined structure was compared to NOE intensity data using complete relaxation matrix back-calculations, yielding an R1x value of 11.2 x 10(-)2. The phenyl ring of the styrene in the S(61,2)C adduct was in the major groove and remained oriented in the 3'-direction as observed for the corresponding S(61,2) adduct paired with thymine [Feng, B., Zhou, L., Pasarelli, M., Harris, C. M., Harris, T. M., and Stone, M. P. (1995) Biochemistry 34, 14021-14036]. A shift of the modified adenine toward the minor groove resulted in the styrenyl ring stacking with nucleotide C5 on the 5'-side of the lesion, which shifted toward the major groove. Unlike the unmodified A.C mismatch, neither the S(61,2)C nor the R(61,2)C adduct formed protonated wobble A.C hydrogen bonds. This suggests that protonated wobble A.C pairing need not be prerequisite to low levels of alpha-SO-induced A --> G mutations. The shift of the modified adenine toward the minor groove in the S(61,2)C structure may play a more important role in the genesis of A --> G mutations. The disordered structure of the R(61,2)C adduct provides a potential explanation as to why that adduct does not induce A --> G mutations.
Painter
S L
SL
Department of Chemistry, Center in Molecular Toxicology, Vanderbilt University, Nashville, Tennessee 37235, USA.
Zegar
I S
IS
Tamura
P J
PJ
Bluhm
S
S
Harris
C M
CM
Harris
T M
TM
Stone
M P
MP
D000225:Adenine / Q000031:analogs & derivatives* / Q000737:chemistry; D020137:Base Pair Mismatch; D003062:Codon / Q000737:chemistry* / Q000235:genetics; D003596:Cytosine / Q000031:analogs & derivatives* / Q000737:chemistry; D018736:DNA Adducts / Q000737:chemistry*; D004852:Epoxy Compounds / Q000737:chemistry*; D011905:Genes, ras; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008958:Models, Molecular; D008968:Molecular Conformation; D019906:Nuclear Magnetic Resonance, Biomolecular; D009838:Oligodeoxyribonucleotides / Q000737:chemistry* / Q000235:genetics; D011522:Protons; D013237:Stereoisomerism; D020058:Styrene / Q000737:chemistry; D013816:Thermodynamics
D003062:Codon; D018736:DNA Adducts; D004852:Epoxy Compounds; D009838:Oligodeoxyribonucleotides; D011522:Protons; C096698:alpha-(N6-adenyl)styrene oxide-DNA adduct; C085540:2'-deoxycytosine; D020058:Styrene; D003596:Cytosine; C013690:styrene oxide; D000225:Adenine