title: Mutagenic Replication of N2-Deoxyguanosine Benzo[a]pyrene Adducts by Escherichia coli DNA Polymerase I and Sulfolobus solfataricus DNA Polymerase IV.
abstract: Benzo[a]pyrene, a potent human carcinogen, is metabolized in vivo to a diol epoxide that reacts with the N2-position of guanine to produce N2-BP-dG adducts. These adducts are mutagenic causing G to T transversions. These adducts block replicative polymerases but can be bypassed by the Y-family translesion synthesis polymerases. The mechanisms by which mutagenic bypass occurs is not well-known. We have evaluated base pairing structures using atomic substitution of the dNTP with two stereoisomers, 2'-deoxy-N-[(7R,8S,9R,10S)-7,8,9,10-tetrahydro-7,8,9-trihydroxybenzo[a]pyren-10-yl]guanosine and 2'-deoxy-N-[(7S,8R,9S,10R)-7,8,9,10-tetrahydro-7,8,9-trihydroxybenzo[a]pyren-10-yl]guanosine. We have examined the kinetics of incorporation of 1-deaza-dATP, 7-deaza-dATP, 2'-deoxyinosine triphosphate, and 7-deaza-dGTP, analogues of dATP and dGTP in which single atoms are changed. Changes in rate will occur if that atom provided a critical interaction in the transition state of the reaction. We examined two polymerases, Escherichia coli DNA polymerase I (Kf) and Sulfolobus solfataricus DNA polymerase IV (Dpo4), as models of a high fidelity and TLS polymerase, respectively. We found that with Kf, substitution of the nitrogens on the Watson-Crick face of the dNTPs resulted in decreased rate of reactions. This result is consistent with a Hoogsteen base pair in which the template N2-BP-dG flipped from the anti to syn conformation. With Dpo4, while the substitution did not affect the rate of reaction, the amplitude of the reaction decreased with all substitutions. This result suggests that Dpo4 bypasses N2-BP-dG via Hoogsteen base pairs but that the flipped nucleotide can be either the dNTP or the template.
authors.affiliation: Department of Biochemistry and Molecular Biology, Milton S. Hershey Medical Center, Pennsylvania State University College of Medicine , Hershey, Pennsylvania 17033, United States.
authors.forename: A S Prakasha
authors.identifier: 0000-0002-0055-7103
authors.initials: ASP
authors.lastname: Gowda
authors.affiliation: Department of Pharmacology, Milton S. Hershey Medical Center, Pennsylvania State University College of Medicine , Hershey, Pennsylvania 17033, United States.
authors.forename: Jacek
authors.initials: J
authors.lastname: Krzeminski
authors.affiliation: Department of Pharmacology, Milton S. Hershey Medical Center, Pennsylvania State University College of Medicine , Hershey, Pennsylvania 17033, United States.
authors.forename: Shantu
authors.initials: S
authors.lastname: Amin
authors.affiliation: Department of Chemistry and Biochemistry, The Ohio State University , Columbus, Ohio 43210, United States.
authors.forename: Zucai
authors.identifier: 0000-0003-3871-3420
authors.initials: Z
authors.lastname: Suo
authors.affiliation: Department of Biochemistry and Molecular Biology, Milton S. Hershey Medical Center, Pennsylvania State University College of Medicine , Hershey, Pennsylvania 17033, United States.
authors.forename: Thomas E
authors.identifier: 0000-0002-6805-3729
authors.initials: TE
authors.lastname: Spratt
mesh_terms: D020029:Base Pairing; D001580:Benzopyrenes / Q000378:metabolism*; D002384:Catalysis; D018736:DNA Adducts; D004256:DNA Polymerase I / Q000378:metabolism*; D019951:DNA Polymerase beta / Q000378:metabolism*; D004261:DNA Replication; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology*; D048229:Sulfolobus solfataricus / Q000201:enzymology*
chemical_list: D001580:Benzopyrenes; D018736:DNA Adducts; C478198:benzo(a)pyrene N2-dG adduct; D004256:DNA Polymerase I; D019951:DNA Polymerase beta; D003849:Deoxyguanosine