Involvement of specialized DNA polymerases in mutagenesis by 8-hydroxy-dGTP in human cells
Involvement of specialized DNA polymerases in mutagenesis by 8-hydroxy-dGTP in human cells
复制标题
DOI：
10.1016/j.dnarep.2008.12.009
复制
发表时间：
2009-05-01
期刊：
影响因子：
3.8
通讯作者：
Kamiya, Hiroyuki
中科院分区：
文献类型：
--
作者：
Satou, Kazuya;Hori, Mika;Kamiya, Hiroyuki
氧化形式的dGTP， 8-羟基-2'-脱氧鸟苷5'-三磷酸（8-OH-dGTP）的诱变性。使用人293T细胞进行检测。首先将含有supF基因的穿梭质粒DNA转染到细胞中，然后通过渗透压法导入8-OH-dGTP。回收细胞中复制的dna，然后将其转染到大肠杆菌中。8-OH-dGTP诱导细胞A:T -> C:G置换突变。sirna对DNA聚合酶eta和zeta以及REV1的敲除减少了A:T -> C:G取代突变，表明这些DNA聚合酶参与了8-OH-dGTP在人类细胞中与A相反的错误掺入。相比之下。DNA聚合酶iota的敲除对8- oh - dgtp诱导的突变没有影响。DNA pol - eta + zeta和REV1 + DNA pol - zeta的双敲低更明显（而DNA pol - eta + REV1的双敲低则不明显），表明REV1-DNA pol - eta和DNA pol - zeta在不同的步骤中起作用。这些结果表明，特殊的DNA聚合酶参与了氧化dGTP诱导的突变。(C) 2008爱思唯尔版权所有。
The mutagenicity of an oxidized form of dGTP, 8-hydroxy-2'-deoxyguanosine 5'-triphosphate (8-OH-dGTP). was examined using human 293T cells. Shuttle plasmid DNA containing the supF gene was first transfected into the cells, and then 8-OH-dGTP was introduced by means of osmotic pressure. The DNAs replicated in the cells were recovered and then transfected into Escherichia coli. 8-OH-dGTP induced A:T -> C:G substitution mutations in the cells. The knock-downs of DNA polymerases eta and zeta, and REV1 by siRNAs reduced the A:T -> C:G substitution mutations, suggesting that these DNA polymerases are involved in the misincorporation of 8-OH-dGTP opposite A in human cells. In contrast. the knock-down of DNA polymerase iota did not affect the 8-OH-dGTP-induced mutations. The decrease in the induced mutation frequency was more evident by double knock-downs of DNA pols eta plus zeta and REV1 plus DNA pol zeta (but not by that of DNA pol eta plus REV1), suggesting that REV1-DNA pol eta and DNA pol zeta work in different steps. These results indicate that specialized DNA polymerases are involved in the mutagenesis induced by the oxidized dGTP. (C) 2008 Elsevier B,V. All rights reserved.