Roles of specialized DNA polymerases in mutagenesis by 8-hydroxyguanine in human cells
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概要
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The formation of 8-hydroxyguanine (8-OH-Gua, 7,8-dihydro-8-oxoguanine) in DNA and in the nucleotide pool results in G:C→T:A and A:T→C:G substitution mutations, respectively, due to the ability of 8-OH-Gua to pair with both C and A. In this study, shuttle plasmid DNAs containing 8-OH-Gua paired with C and A in the supF gene were transfected into human 293T cells, in which specialized DNA polymerases were knocked-down. The DNAs replicated in the cells were recovered and then introduced into an indicator strain of Escherichia coli. Mutation analysis indicated that the knock-downs of DNA polymerases η and ζ by siRNAs enhanced the G:C→T:A mutations caused by 8-OH-Gua:C. The 8-OH-Gua:C-induced mutation frequency was not further increased by double knock-downs of DNA polymerases η and ζ, suggesting that the two DNA polymerases work in the same pathway. In addition, the reduction of DNA polymerase η slightly decreased the A:T→C:G substitutions caused by 8-OH-Gua:A. These results suggest that DNA polymerases η and ζ are involved in the bypass of 8-OH-Gua in human cells.
- Elsevier B.V.の論文
- 2010-04-01
Elsevier B.V. | 論文
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