Studies on the function of Werner syndrome gene product and analyses of the mechanism to induce aging related symptoms

维尔纳综合征基因产物的功能研究及衰老相关症状的机制分析

• 批准号: 18390019
• 负责人: ENOMOTO Takemi
• 金额: $ 10.7万
• 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: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18390019/
• 关键词: gene; cancer; aging; Werner syndrome; WRN; reactive oxigen; SOD1; SOD2; 糖尿病

The aim of this study is to clarify the mechanism to cause predisposition of cancer due to genome instability and many aging related symptoms including type II diabetes mellitus in Werner syndrome patients by investigating the functions of WRN in DNA replication and repair. In this study, we used yeast model system to know the functional relationship between WRN, WRNIP1 and DNA polymerase δ (Polδ), and various gene disrupted chicken DT40 cells to analyze the pathway in which WRN functions. In addition, we performed biochemical analyses using purified enzymes and proteins. We isolated many mutants having mutations in the subunit of Polδ, pol31 and got evidence indicating functional relationship between WRN, WRNIP1 and pol31. In addition, by using many DT40 mutants it was suggested that WRN functions with RAD52 and in the pathway in which RAD18 is involved when cells are caused specific DNA lesions. Furthermore, it was suggested that WRNIP1 functions upstream of RAD18 and directly interacts with RAD18. Finally, we established the base to know the function of WRN under oxidative stress by constructing SOD1 or SOD2 gene defective mutants using DT40 cells.

本研究的目的是通过研究WRN在DNA复制和修复中的功能，阐明Werner综合征患者因基因组不稳定性和许多与衰老相关的症状（包括II型糖尿病）而导致癌症易感性的机制。本研究利用酵母模型系统研究了WRN、WRNIP1和DNA聚合酶δ（Polδ）之间的功能关系，并通过对鸡DT 40细胞进行不同基因的破坏，分析了WRN的功能途径。此外，我们使用纯化的酶和蛋白质进行了生化分析。我们分离了许多在Polδ、pol 31亚基上发生突变的突变体，并获得了表明WRN、WRNIP 1和pol 31之间功能关系的证据。此外，通过使用许多DT 40突变体，表明WRN与RAD 52一起起作用，并且当细胞引起特异性DNA损伤时，WRN在其中涉及RAD 18的途径中起作用。此外，有人建议WRNIP1的功能上游的RAD18和直接与RAD18相互作用。最后，我们利用DT 40细胞构建了SOD 1或SOD 2基因缺陷突变体，为了解WRN在氧化应激下的功能奠定了基础。

项目成果

Analyses of functional interaction between RECQL1, RECQL5, and BLM which physically interact with DNA topoisomerase IIIa

分析 RECQL1、RECQL5 和 BLM 之间的功能相互作用，这些相互作用与 DNA 拓扑异构酶 IIIa 发生物理相互作用

• 发表时间: 2008
• 期刊: Biochim. Biophys. Acta. 1782
• 作者: Otsuki M;Seki M;Inoue E;Abe T;Narita Y;Yoshimura A;Tada S;Ishii Y;Enomoto T
• 通讯作者: Enomoto T

Mgsl and Rad l8/Rad5/Mms2 are required for survival of novel temperature/cold sensitive mutant alleles of Po131, the second subunit of DNA polymerase δ, in Sacharomyces cerevisiae.

Mgs1和Rad18/Rad5/Mms2是酿酒酵母中DNA聚合酶δ的第二个亚基Po131的新型温度/冷敏感突变等位基因的存活所必需的。

• 发表时间: 2006
• 期刊: DNA Repair 5・12
• 作者: Takeya;R.;Ueno;N.;and Sumimoto;H.;Akari Yoshimura;成相裕子;横溝岳彦;N.Davoodi Vijeh Motlagh
• 通讯作者: N.Davoodi Vijeh Motlagh

Dpb11, the budding yeast homologue of TopBP1, functions with the checkpoint clamp in recombination repair

Dpb11 是 TopBP1 的出芽酵母同源物，在重组修复中与检查点钳一起发挥作用

• 发表时间: 2006
• 期刊: Nucleic Acids Res. 34
• 作者: Naoe T;Suzuki T;Kiyoi H and Urano T;Hideaki Ogiwara
• 通讯作者: Hideaki Ogiwara

Rad52 sumoylation and its involvement in the efficient induction of homologous recombination

• DOI: 10.1016/j.dnarep.2008.02.005
• 发表时间: 2008-06-01
• 期刊: DNA REPAIR
• 影响因子: 3.8
• 作者: Ohuchi, Takashi;Seki, Masayuki;Enomoto, Takemi
• 通讯作者: Enomoto, Takemi

A novel Rad18 function involved in the protection of the vertebrate genome after exposure to camptothecin

Rad18 的一种新功能参与暴露于喜树碱后脊椎动物基因组的保护

• 发表时间: 2006
• 期刊: DNA Repair 5
• 作者: Yoshimura;A.;Nishino;K.;Takezawa;J.;Tada;S.;Kobayashi;T.;Sonoda;E.;Kawamoto;T.;Takeda;S.;Ishii;Y.;Yamada;K.;Enomoto;T.;Seki;M
• 通讯作者: M