Mutagenesis of benzo[a]pyrene diol epoxide in yeast: Requirement for DNA polymerase ζ and involvement of DNA polymerase η

Zhongwen Xie, Elena Braithwaite, Dongyu Guo, Bo Zhao, Nicholas Geacintov, Zhigang Wang

Research output: Contribution to journalArticle

Abstract

Benzo[a]pyrene is a potent environmental carcinogen, which can be metabolized in cells to the DNA damaging agent anti-benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide (anti-BPDE). We hypothesize that mutations induced by BPDE DNA adducts are mainly generated through an error-prone translesion synthesis that requires a specialized DNA polymerase (Pol). Using an in vivo mutagenesis assay in the yeast model system, we have examined the potential roles of Polζ and Polη in (± )-anti-BPDE-induced mutagenesis. In cells proficient in mutagenesis, (±)-anti-BPDE induced 85% base substitutions with predominant G → C followed by G → T transversions, 9% deletions of 1-3 nucleotides, and 6% insertions of 1-3 nucleotides. In rad30 mutant cells lacking Polζ, (±)-anti-BPDE-induced mutagenesis was reduced and accompanied by a moderate decrease in base substitutions and more significant decrease in deletions and insertions of 1-3 nucleotides. In rev3 mutant cells Polζ, (±)-anti-BPDE-induced mutagenesis was mostly abolished, leading to a great decrease in both base substitutions and deletions/insertions of 1-3 nucleotides. In contrast, large deletions/insertions were significantly increased in cells lacking Polζ. Consistent with the in vivo results, purified yeast Polζ performed limited translesion synthesis opposite (+)- and (-)-trans-anti-BPDE-N2-dG DNA adducts with predominant G incorporation opposite the lesion. These results show that (± )-anti-BPDE-induced mutagenesis in yeast requires Polζ and partially involves Polη and suggest that Polζ directly participates in nucleotide insertions opposite the lesion, while Polη significantly contributes to deletions and insertions of 1-3 nucleotides.

Original languageEnglish (US)
Pages (from-to)11253-11262
Number of pages10
JournalBiochemistry
Volume42
Issue number38
DOIs
StatePublished - Sep 30 2003

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7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide
Mutagenesis
Benzo(a)pyrene
Epoxy Compounds
DNA-Directed DNA Polymerase
Yeast
Yeasts
Nucleotides
Substitution reactions
DNA Adducts
Environmental Carcinogens
Assays
Mutation
DNA

ASJC Scopus subject areas

  • Biochemistry

Cite this

Mutagenesis of benzo[a]pyrene diol epoxide in yeast : Requirement for DNA polymerase ζ and involvement of DNA polymerase η. / Xie, Zhongwen; Braithwaite, Elena; Guo, Dongyu; Zhao, Bo; Geacintov, Nicholas; Wang, Zhigang.

In: Biochemistry, Vol. 42, No. 38, 30.09.2003, p. 11253-11262.

Research output: Contribution to journalArticle

Xie, Zhongwen ; Braithwaite, Elena ; Guo, Dongyu ; Zhao, Bo ; Geacintov, Nicholas ; Wang, Zhigang. / Mutagenesis of benzo[a]pyrene diol epoxide in yeast : Requirement for DNA polymerase ζ and involvement of DNA polymerase η. In: Biochemistry. 2003 ; Vol. 42, No. 38. pp. 11253-11262.
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