Direct synthesis and indentification of benzo[a]pyrene diol epoxide-deoxyguanosine binding sites in modified oligodeoxynucleotides

Journal Article · · Chemical Research in Toxicology; (United States)

OSTI ID:6282418

Mao, B; Margulis, L A; Li, B; Ibanez, V; Geacintov, N E ^[1]; Harvey, R G; Lee, H ^[2]

---

1. New York Univ., NY (United States)
2. Univ. of Chicago, IL (United States)

Adducts derived from the reaction of the benzo[a]pyrene metabolite model compound (+)-anti-7[beta], 8[alpha]-dihydroxy-9[alpha],10[alpha]-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene [(+)-BPDE] with the single-stranded oligodeoxynucleotide 5'-d(TATGCGTAT) were obtained according to direct synthesis techniques described earlier [Cosman, M., Ibanez, V., Geacintov, N.E., and Harvey, R.G.(1990) Carcinogenesis 11,1667-1672]. Four major adducts, involving trans and cis addition (trans/cis adduct ratio [approx] 4.5) of (+)-BPDE to the exocyclic amino groups of guanines G[sub 4] and G[sub 6] (the numbers denote the positions of the guanines counted from the 5'-side) were obtained. These adducts can be separated from one another by reverse-phase high-performance liquid chromatography methods. The site of BPDE binding on either G[sub 4] or G[sub 6] can be determined from the electrophoresis band patterns on 20% polyacrylamide gels of the BPDE-modified oligonucleotides subjected to the G+A and G Maxam-Gilbert strand cleavage reactions [Maxam, A.M., and Gilbert, W. (1980) Methods. Enzymol. 65, 499-560]. The electrophoresis gel band patterns are different for unmodified DNA and the two different BPDE residues migrate slower than the corresponding fragments derived from the unmodified oligonucleotide and (2) strand cleavage tends to be inhibited on the 5'-sides of BPDE-modified guanines in the G+A, but not the G reaction.

• DOE Contract Number: FG02-88ER60674; FG02-86ER60405
• OSTI ID: 6282418
• Journal Information: Chemical Research in Toxicology; (United States), Journal Name: Chemical Research in Toxicology; (United States) Vol. 5:6; ISSN 0893-228X; ISSN CRTOEC
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
550300 -- Cytology
59 BASIC BIOLOGICAL SCIENCES
ADDUCTS
AMINES
AROMATICS
AZAARENES
BIOCHEMICAL REACTION KINETICS
CARCINOGENESIS
CHROMATOGRAPHY
COLLOIDS
DISPERSIONS
DNA
DNA ADDUCTS
ELECTROPHORESIS
GELS
GUANINE
HETEROCYCLIC COMPOUNDS
HYDROCARBONS
HYDROXY COMPOUNDS
KINETICS
NUCLEIC ACIDS
OLIGONUCLEOTIDES
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PATHOGENESIS
POLYCYCLIC AROMATIC HYDROCARBONS
PURINES
REACTION KINETICS
SEPARATION PROCESSES
STRAND BREAKS
SYNTHESIS