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Molecular mechanism to couple DNA replication with avoidance of DNA lesions, the defect in which induces cancer and aging

DNA复制与避免DNA损伤相结合的分子机制，DNA损伤会诱发癌症和衰老

基本信息

批准号：
15390021
负责人：
ENOMOTO Takemi
金额：
$ 9.73万
依托单位：
Tohoku University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15390021/
关键词：
Werner syndrome progeria WRN WRNIP1 RecQ family helicase DNA polymerase δ homologous recombination 複製後修復

项目摘要

The aim of this study is to clarify the molecular mechanism to couple DNA replication with the avoidance of DNA lesions by analyzing the processes in which RecQ family helicases and WRNIP1 (Werner helicase interacting protein 1) are involved and to elucidate the mechanism to induce cancer and aging due to the defect in the coupling.The analyses using yeast indicated that Wrnip1 is functionally related to DNA polymerase δ (Polδ), PCNA, and RFC and interacts directly with Polδ. In addition, by using many mutants having mutations in the subunit of Polδ, pol31, which we isolated, it was revealed that Wrnip1 and Sgs1 (WRN homologue in yeast) play very important roles when Polδ has some defects. Biochemical analysis using purified human WRNIP1, Polδ, PCNA, and RFC indicated that WRNIP1 binds to Polδ and stimulates its activity. The mechanism of the stimulation was the enhancement of initiation frequency of DNA synthesis. To analyze the pathways in which WRN and WRNIP1 are involved, we constructed various gene knockout cells using chicken DT40 cells. The analyses using these cells suggested that WRN and WRNIP1 function in different repair pathways, and WRN functions in a recombination repair pathway in which Rad52 is involved but Xrcc3 is not. Furthermore, it was suggested that WRN somehow functionally interacts with Ku70, which is involved in nonhomologous end-joining.

本研究的目的是通过分析RecQ家族解旋酶和WRNIP1在DNA复制过程中的作用，阐明DNA复制与避免DNA损伤的分子机制。利用酵母细胞进行的分析表明，Werner nip1与DNA聚合酶δ（Polδ）在功能上相关，PCNA和RFC，并直接与Polδ相互作用。另外，通过对我们分离的Polδ亚基pol31突变株的研究，发现当Polδ发生缺陷时，pnip 1和Sgs 1（WRN同源物）起着重要作用。使用纯化的人WRNIP1、Polδ、PCNA和RFC进行的生化分析表明，WRNIP1与Polδ结合并刺激其活性。其作用机制是提高DNA合成的起始频率。为了分析WRN和WRNIP1参与的途径，我们使用鸡DT 40细胞构建了各种基因敲除细胞。使用这些细胞的分析表明，WRN和WRNIP1在不同的修复途径中起作用，WRN在重组修复途径中起作用，其中Rad52参与，但Xrcc 3不参与。此外，研究表明WRN在某种程度上与Ku70在功能上相互作用，而Ku70参与非同源末端连接。

项目成果

期刊论文数量（64）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

SMC6 is required for MMS-induced interchromosomal and sister chromatid recombinations in Saccharomyces cerevisiae.

DOI：
10.1016/j.dnarep.2003.12.007
发表时间：
2004-04
期刊：
DNA repair
影响因子：
3.8
作者：
F. Onoda;M. Takeda;M. Seki;D. Maeda;J. Tajima;A. Ui;H. Yagi;T. Enomoto
通讯作者：
F. Onoda;M. Takeda;M. Seki;D. Maeda;J. Tajima;A. Ui;H. Yagi;T. Enomoto

Molecular mechanism of Werner's syndrome

维尔纳综合征的分子机制

DOI：
发表时间：
2004
期刊：
影响因子：
0
作者：
Seiki Hirano;Ayako Ui;Seiki Hirano;Ayako Ui;Changwok Lee;Fumitoshi Onoda;Wensheng Wang;Changwok Lee;Fumitoshi Onoda;Wensheng Wang;Daisuke Maeda;Wensheng Wang;Wensheng Wang;Dana Branzei;Dana Branzei;Dana Branzei;Dana Branzei;Hirobumi Suzuki;Ayako Ui;Hirobumi Suzuki;Ayako Ui;Dana Branzei
通讯作者：
Dana Branzei

Fumitoshi Onoda: "SMC6 is required for MMS-induced interchromosomal and sister chromatid recombinations in Saccharomyces cerevisiae"DNA Repair. (in press).

Fumitoshi Onoda：“SMC6 是酿酒酵母中 MMS 诱导的染色体间和姐妹染色单体重组所必需的”DNA 修复。