trans-lesion synthesis past bulky benzo[a]pyrene diol epoxide N2-dG and N6-dA lesions catalyzed by DNA bypass polymerases

Olga Rechkoblit, Yanbin Zhang, Dongyu Guo, Zhigang Wang, Shantu Amin, Jacek Krzeminsky, Natalia Louneva, Nicholas Geacintov

Research output: Contribution to journalArticle

Abstract

The effectiveness of in vitro primer elongation reactions catalyzed by human bypass DNA polymerases κ (hDinB1), pol η (hRad30A), pol ι (hRad30B), and yeast pol ζ (Rev3 and Rev7) in site-specifically modified template oligonucleotide strands were studied in vitro. The templates contained single bulky lesions derived from the trans-addition of the mutagenic (+)- or (-)-enantiomers of r7,t8-dihydroxy-t9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene (a metabolite of the environmental carcinogen benzo[a]pyrene), to the exocyclic amino groups of guanine or adenine in oligonucleotide templates 33, or more, bases long. In "running start" primer extension reactions, pol κ effectively bypassed both the stereoisomeric (+)- and (-)-trans-guanine adducts but not the analogous adenine adducts. In sharp contrast, pol η, which exhibits considerable sequence homology with pol κ (both belong to the group of Y family polymerases), is partially blocked by the guanine adducts and the (-)-trans-adenine adduct, although the stereoisomeric (+)-trans-adenine adduct is more successfully bypassed. Neither pol ι nor pol ζ, either alone or in combination, were effective in trans-lesion synthesis past the same adducts. In all cases, the fidelity of insertion is dependent on adduct stereochemistry and structure. Generally, error-free nucleotide insertion opposite the lesions tends to depend more on adduct stereochemistry than error-prone insertion. None of the polymerases tested are a universal bypass polymerase for the stereoisomeric bulky polycyclic aromatic hydrocarbon-DNA adducts derived from anti-BPDE.

Original languageEnglish (US)
Pages (from-to)30488-30494
Number of pages7
JournalJournal of Biological Chemistry
Volume277
Issue number34
DOIs
StatePublished - Aug 23 2002

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Benzo(a)pyrene
Epoxy Compounds
Adenine
DNA-Directed DNA Polymerase
Guanine
Stereochemistry
DNA
Oligonucleotides
7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide
Environmental Carcinogens
Enantiomers
Sequence Homology
Metabolites
Running
Yeast
Elongation
Nucleotides
Yeasts
In Vitro Techniques

ASJC Scopus subject areas

  • Biochemistry

Cite this

trans-lesion synthesis past bulky benzo[a]pyrene diol epoxide N2-dG and N6-dA lesions catalyzed by DNA bypass polymerases. / Rechkoblit, Olga; Zhang, Yanbin; Guo, Dongyu; Wang, Zhigang; Amin, Shantu; Krzeminsky, Jacek; Louneva, Natalia; Geacintov, Nicholas.

In: Journal of Biological Chemistry, Vol. 277, No. 34, 23.08.2002, p. 30488-30494.

Research output: Contribution to journalArticle

Rechkoblit, Olga ; Zhang, Yanbin ; Guo, Dongyu ; Wang, Zhigang ; Amin, Shantu ; Krzeminsky, Jacek ; Louneva, Natalia ; Geacintov, Nicholas. / trans-lesion synthesis past bulky benzo[a]pyrene diol epoxide N2-dG and N6-dA lesions catalyzed by DNA bypass polymerases. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 34. pp. 30488-30494.
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T1 - trans-lesion synthesis past bulky benzo[a]pyrene diol epoxide N2-dG and N6-dA lesions catalyzed by DNA bypass polymerases

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AU - Zhang, Yanbin

AU - Guo, Dongyu

AU - Wang, Zhigang

AU - Amin, Shantu

AU - Krzeminsky, Jacek

AU - Louneva, Natalia

AU - Geacintov, Nicholas

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