Translesion synthesis by human DNA polymerase κ on a DNA template containing a single stereoisomer of dG-(+)- or dG-(-)-anti-N2-BPDE (7,8-dihydroxy-anti-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene)

Naomi Suzuki, Eiji Ohashi, Alexander Kolbanovskiy, Nicholas E. Geacintov, Arthur P. Grollman, Haruo Ohmori, Shinya Shibutani

研究成果: Contribution to journalArticle査読

143 被引用数 (Scopus)

抄録

Several recently discovered human DNA polymerases are associated with translesion synthesis past DNA adducts. These include human DNA polymerase κ (pol κ), a homologue of Escherichia coli pol IV, which enhances the frequency of spontaneous mutation. Using a truncated form of κ (pol κδC), translesion synthesis past dG-(+)- or dG-(-)-anti-N2-BPDE (7,8-dihydroxy-anti-9,10-epoxy-7,8,9,-10-tetrahydrobenzo[a]pyrene) adducts was explored. Site-specifically-modified oligodeoxynucleotides containing a single stereoisomeric dG-N2-BPDE lesion were used as DNA templates for primer extension reactions catalyzed by pol κ δC. Primer extension was retarded one base prior to the dG-N2-BPDE lesion; when incubated for longer times or with higher concentration of enzyme, full primer extension was observed. Quantitative analysis of fully extended products showed preferential incorporation of dCMP, the correct base, opposite all four stereoisomeric dG-N2-BPDE lesions. (+)-trans-dG-N2-BPDE, a major BPDE-DNA adduct, promoted small amounts of dTMP, dAMP, and dGMP misincorporation opposite the lesion (total 2.7% of the starting primers) and deletions (1.1%). Although (+)-cis-dG-N2-BPDE was most effective in blocking translesion synthesis, its miscoding properties were similar to other dG-N2-BPDE isomers. Steady-state kinetic data indicate that dCMP is efficiently inserted opposite all dG-N2-BPDE adducts and extended past these lesions. The relative frequency of translesion synthesis (Fins x Fext) of dC·dG-N2-BPDE pairs was 2-6 orders of magnitude higher than that of other mismatched pairs. Pol κ may play an important role in translesion synthesis by incorporating preferentially the correct base opposite dG-N2-BPDE. Its relatively low contribution to mutagenicity suggests that other newly discovered DNA polymerase(s) may be involved in mutagenic events attributed to dG-N2-BPDE adducts in human cells.

本文言語英語
ページ(範囲)6100-6106
ページ数7
ジャーナルBiochemistry
41
19
DOI
出版ステータス出版済み - 5 14 2002
外部発表はい

All Science Journal Classification (ASJC) codes

  • 生化学

フィンガープリント

「Translesion synthesis by human DNA polymerase κ on a DNA template containing a single stereoisomer of dG-(+)- or dG-(-)-anti-N<sup>2</sup>-BPDE (7,8-dihydroxy-anti-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene)」の研究トピックを掘り下げます。これらがまとまってユニークなフィンガープリントを構成します。

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