Identification of pathways controlling DNA damage induced mutation in Saccharomyces cerevisiae
Identification of pathways controlling DNA damage induced mutation in Saccharomyces cerevisiae
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DOI：
10.1016/j.dnarep.2008.02.007
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发表时间：
2008-05-03
期刊：
影响因子：
3.8
通讯作者：
Romesberg, Floyd E.
中科院分区：
文献类型：
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作者：
Lis, Ewa T.;O'Neill, Bryan M.;Romesberg, Floyd E.
大多数类型的 DNA 损伤引起的突变被认为是由复制后修复的易错分支和相关的跨损伤合成聚合酶介导的。为了进一步了解 DNA 损伤的诱变反应，我们筛选了 4848 个酿酒酵母单倍体基因缺失菌株，以减少损伤诱导的 CAN1 基因突变。通过对筛选鉴定的菌株进行广泛的定量验证，我们鉴定了 10 个基因，其中包括已知参与诱导突变的容易出错的复制后修复基因，以及另外两个基因 FYV6 和 RNR4。我们证明 FYV6 和 RNR4 在诱导突变方面是上位的，并且它们的功能至少部分独立于复制后修复。这种诱导突变的途径似乎是由 dNTP 水平的增加介导的，dNTP 水平的增加有助于复制聚合酶 Pol δ 绕过病变，并且在 UV 和 MMS 诱导的突变的情况下，它与容易出错的复制后修复一样重要，但仅负责 EMS 诱导的突变。我们表明，羟基脲（一种核糖核苷酸还原酶的小分子抑制剂）可以有效抑制 Rnr4/Pol δ 诱导的突变，这表明如果人类细胞中存在类似的途径，则可能可以干预某些形式的突变。 (c) 2008 Elsevier B.V. 保留所有权利。
Mutation in response to most types of DNA damage is thought to be mediated by the error-prone sub-branch of post-replication repair and the associated translesion synthesis polymerases. To further understand the mutagenic response to DNA damage, we screened a collection of 4848 haploid gene deletion strains of Saccharomyces cerevisiae for decreased damage-induced mutation of the CAN1 gene. Through extensive quantitative validation of the strains identified by the screen, we identified ten genes, which included error-prone post-replication repair genes known to be involved in induced mutation, as well as two additional genes, FYV6 and RNR4. We demonstrate that FYV6 and RNR4 are epistatic with respect to induced mutation, and that they function, at least partially, independently ofpostreplication repair. This pathway of induced mutation appears to be mediated by an increase in dNTP levels that facilitates lesion bypass by the replicative polymerase Pol delta, and it is as important as error-prone post-replication repair in the case of UV- and MMS-induced mutation, but solely responsible for EMS-induced mutation. We show that Rnr4/Pol delta-induced mutation is efficiently inhibited by hydroxyurea, a small molecule inhibitor of ribonucleotide reductase, suggesting that if similar pathways exist in human cells, intervention in some forms of mutation may be possible. (c) 2008 Elsevier B.V. All rights reserved.