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

• 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

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.

澄清分子机制的目的是涉及ECQ家族的螺旋量和WRNIP1（Werner Helicase相互作用蛋白1），并阐明了使用FC的分析中的缺陷诱导癌症和衰老，通过在我们隔离的Polδ的亚基中使用许多MUT突变，可以透露WRNIP1和SGS1（年度为WRN同源物。使用纯化的人WRNIP1，POLΔ，PCNA，PCNA和RFC的ICHEMITAL分析表明WRNIP1与POLΔ与PolΔ结合并刺激DNA合成的娱乐性。在RAD52的重组修复途径中，建议WRN在功能上与KU70相互作用，而Ku70与非综合学的最终连接有关。

项目成果

Caenorhabditis elegans geminin homologue particpates in cell cycle regulation and germline development.

秀丽隐杆线虫双子蛋白同源物参与细胞周期调节和种系发育。

• 发表时间: 2005
• 期刊: Journal of Biological Chemistry (in press)
• 作者: Yanagi;K.;et al.
• 通讯作者: et al.

Sevim Isik: "The SUMO pathway is required for selective degradation of DNA topoisomerase IIβ induced by a catalytic inhibitor ICRF-193"FEBS Letters. 546. 374-378 (2003)

Sevim Isik：“催化抑制剂 ICRF-193 诱导的 DNA 拓扑异构酶 IIβ 的选择性降解需要 SUMO 途径”FEBS Letters 546. 374-378 (2003)。

U6c9 is required for damage-tolerance and damage-induced interchromosomal homologue recombination in Saccharomyces cerevisiae.

U6c9 是酿酒酵母中损伤耐受和损伤诱导的染色体间同源重组所必需的。

• 发表时间: 2004
• 期刊: DNA Repair 3
• 作者: Yoshihito Ueno;Daisuke Maeda
• 通讯作者: Daisuke Maeda

Nao Odagiri: "Budding yeast mms4 is epistatic with rad52 and its function can be replaced by a bacterial Holliday junction resolvase"DNA repair. 2. 347-358 (2003)

Nao Odagiri：“芽殖酵母 mms4 与 rad52 上位，其功能可以被细菌霍利迪连接体解析酶取代”DNA 修复。

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