Basic study on the mechanisms of carcinogenesis and aging with special reference to the function of RecQ family helicases

致癌和衰老机制的基础研究，特别是RecQ家族解旋酶的功能

• 批准号: 11307055
• 负责人: ENOMOTO Takemi
• 金额: $ 24.75万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11307055/
• 关键词: RecQ family; Helicase; Werner syndrome; WHIP; WRN; carcinogenesis; aging; yeast Sgs1; RecQL1; ワーナー症候群; Whip; ブルーム症候群

In this study, we tried to clarify the function of RecQ family helicases to understand the genomic instability resulting in predisposition to cancer and premature aging due to the defect in RecQ family helicases, and obtained following results by using budding yeasts and chicken DT40 cells, which are easily applicable to genetic analysis, as well as cultured human cells.1. It has been indicated by generating and analyzing BLM-/- and BLM-/-/RAD54-/- DT40 cells that the majority of increased SCEs in BLM disrupted cells are formed via homologous recombination and BLM functions to decrease the formation of DNA double strand breaks during DNA replication.2. Werner syndrome gene product, WRN, interacted with Ubc9 and SUMO-1 and was actually conjugated with SUMO-1.3. A novel Werner helicase interacting protein, WHIP, which we found, was co-localized with WRN in the nuclei, and the interaction of these proteins required ATP, indicating a dynamic interaction of these proteins.4. The genetic analyzes using yeast mutants that overexpression of WHIP in mutants of DNA polymerase δ, RFC, or PCNA resulted in the lethality of the cells indicated the existence of functional relationship between WHIP and DNA polymerase δ, RFC, and PCNA. In addition, physical interaction between WHIP and DNA polymerase δ was indicated by the yeast-two hybrid system.5. Analyzes using yeasts also indicated the genetical interaction of the yeast BLM/WRN homologue, Sgs1 with Mre11, Sld3, and Sld5 which are involved in the initiation of DNA replication.

在这项研究中，我们试图了解RECQ的功能，以了解导致癌症衰老预测的基因组不稳定性，因此很容易适用于遗传分析4 - / - DT40细胞，大多数细胞是大多数细胞BLM中断的细胞中的SCE增加是通过同源重组形成的，以减少ENE产物的形成，WRN，与UBC9和Sumo-1相互作用，实际上是与SUMO-1.3相互轭的。与核共定位，蛋白质的相互作用需要蛋白质的ATP IC相互作用。4。 whip和DNAδ，RFC和PCNA的存在。此外，whip和DNA聚合酶δ之间的物理相互作用是由YearS -Two Hybrid System表示的。5 ，SLD3，SLD3 D5参与DNA复制的启动。

项目成果

Tamae Kawabe: "Differential regulation of human RecQ family helicases in cell transformation and cell cycle"Oncogene. 19. 4764-4772 (2001)

Tamae Kawabe：“人类 RecQ 家族解旋酶在细胞转化和细胞周期中的差异调节”癌基因。

Takemi Enomoto: "Function of RecQ family helicases : possible involvement of Bloom's and Werner's syndrome gene products in guarding genome integrity during DNA replication"Journal of Biochemistry. 129. 501-507 (2001)

Takemi Enomoto：“RecQ 家族解旋酶的功能：布卢姆综合征和沃纳综合征基因产物可能参与 DNA 复制过程中保护基因组完整性”《生物化学杂志》。

Atsuko Miyajima: "Sgs1 helicase activity is required for mitotic but apparently not for meiotic functions."Molecular and Cellular Biology. 20. 6399-6409 (2000)

Atsuko Miyajima：“Sgs1 解旋酶活性是有丝分裂所必需的，但显然不是减数分裂功能所必需的。”分子和细胞生物学。

Tamae Kawabe: "Differential regulation of human RecQ family helicases in cell transformation and cell cycle."Oncogene. 19. 4764-4772 (2000)

Tamae Kawabe：“人类 RecQ 家族解旋酶在细胞转化和细胞周期中的差异调节。”癌基因。

Ayako Ui: "Possible involvement of Top3 via the N-terminal region of Sgs1 in the complementation of MMS sensitivity and the suppression of hyper recombination"Mol. Gen. Genet.. 265. 837-850 (2001)

Ayako Ui：“Top3 可能通过 Sgs1 的 N 末端区域参与 MMS 敏感性的互补和超重组的抑制”Mol。