Eukaryotic Y-family polymerases bypass a 3-methyl-2'-deoxyadenosine analog in vitro and methyl methanesulfonate-induced DNA damage in vivo.
Eukaryotic Y-family polymerases bypass a 3-methyl-2'-deoxyadenosine analog in vitro and methyl methanesulfonate-induced DNA damage in vivo.
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DOI：
10.1093/nar/gkn058
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发表时间：
2008-04
影响因子：
14.9
通讯作者：
Woodgate R
中科院分区：
文献类型：
--
作者：
Plosky BS;Frank EG;Berry DA;Vennall GP;McDonald JP;Woodgate R
N3-甲基腺嘌呤（3 MeA）是SN 2甲基化剂在DNA中形成的主要细胞毒性损伤。该病变可能会阻止细胞复制酶的进展，因为N3-甲基阻碍了聚合酶和DNA小沟之间的相互作用。然而，这一假设尚未得到严格证明，因为3 MeA本质上是不稳定的，并且转化为脱碱基位点，而脱碱基位点本身就是一种阻断病变。为了避免这些问题，我们已经化学合成了一个3-deaza类似物的3 MeA（3dMeA）作为一个稳定的亚磷酰胺，并已纳入类似物合成寡核苷酸已在体外作为模板用于DNA复制。正如预期的那样，3dMeA损伤阻断了人类DNA聚合酶α和δ。相比之下，人聚合酶η、1和κ以及酿酒酵母polη能够绕过病变，尽管具有不同的效率和准确性。为了证实我们的研究结果的生理相关性，我们表明，在S。在缺乏Mag 1依赖性3 MeA修复的酿酒酵母中，polη（Rad 30）有助于暴露于甲磺酸甲酯（MMS）的细胞的存活，并且在不存在Mag 1、Rad 30和Rev 3的情况下，人聚合酶η、I和κ能够恢复正常MMS敏感菌株的MMS抗性。
N3-methyl-adenine (3MeA) is the major cytotoxic lesion formed in DNA by SN2 methylating agents. The lesion presumably blocks progression of cellular replicases because the N3-methyl group hinders interactions between the polymerase and the minor groove of DNA. However, this hypothesis has yet to be rigorously proven, as 3MeA is intrinsically unstable and is converted to an abasic site, which itself is a blocking lesion. To circumvent these problems, we have chemically synthesized a 3-deaza analog of 3MeA (3dMeA) as a stable phosphoramidite and have incorporated the analog into synthetic oligonucleotides that have been used in vitro as templates for DNA replication. As expected, the 3dMeA lesion blocked both human DNA polymerases α and δ. In contrast, human polymerases η, ι and κ, as well as Saccharomyces cerevisiae polη were able to bypass the lesion, albeit with varying efficiencies and accuracy. To confirm the physiological relevance of our findings, we show that in S. cerevisiae lacking Mag1-dependent 3MeA repair, polη (Rad30) contributes to the survival of cells exposed to methyl methanesulfonate (MMS) and in the absence of Mag1, Rad30 and Rev3, human polymerases η, ι and κ are capable of restoring MMS-resistance to the normally MMS-sensitive strain.
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影响因子：
2.9
作者：
Monti, P;Iannone, R;Fronza, G
通讯作者：
Fronza, G
影响因子：
4.8
作者：
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通讯作者：
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11.4
作者：
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10.5
作者：
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3.2
作者：
MORRISON, A;CHRISTENSEN, RB;LAWRENCE, CW
通讯作者：
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