WEKO3
アイテム
Dependence of DNA double strand break repair pathways on cell cycle phase in human lymphoblastoid cells
https://repo.qst.go.jp/records/46023
https://repo.qst.go.jp/records/46023e8f537f9-1236-41d8-9146-b331d524c9a4
| Item type | 学術雑誌論文 / Journal Article(1) | |||||
|---|---|---|---|---|---|---|
| 公開日 | 2011-03-30 | |||||
| タイトル | ||||||
| タイトル | Dependence of DNA double strand break repair pathways on cell cycle phase in human lymphoblastoid cells | |||||
| 言語 | ||||||
| 言語 | eng | |||||
| 資源タイプ | ||||||
| 資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||
| 資源タイプ | journal article | |||||
| アクセス権 | ||||||
| アクセス権 | metadata only access | |||||
| アクセス権URI | http://purl.org/coar/access_right/c_14cb | |||||
| 著者 |
Takashima, Yoshio
× Takashima, Yoshio× Honma, Masamitsu× et.al× 高島 良生 |
|||||
| 抄録 | ||||||
| 内容記述タイプ | Abstract | |||||
| 内容記述 | DNA double-strand breaks (DSBs) are usually repaired by nonhomologous end-joining (NHEJ) or homologous recombination (HR). NHEJ is thought to be the predominant pathway operating in mammalian cells functioning in all phases of the cell cycle, while HR works in the late-S and G2 phases. However, relative contribution, competition, and dependence on cell cycle phases are not fully understood. We previously developed a system to trace the fate of DSBs in the human genome by introducing the homing endonuclease I-SceI site into the thymidine kinase (TK) gene of human lymphoblastoid TK6 cells. Here, we use this system to investigate the relative contribution of HR and NHEJ for repairing I-SceI-induced DSBs under various conditions. We used a novel transfection system, AmaxaTM nucleofector, which directly introduces the I-SceI expression vector into cell nuclei. Approximately 65% of transfected cells expressed the I-SceI enzyme and over 50% of the cells produced a single DSB in the genome. The relative contribution of NHEJ and HR for repairing the DSB was 100:1 and did not change with transfection efficiency. Cotransfection with KU80-siRNA significantly diminished KU80 protein levels and decreased NHEJ activity, but did not increase HR. We also investigated HR and NHEJ in synchronized cells. The HR frequency was 2-3 times higher in late-S/G2 phases than in G1, whereas NHEJ was unaffected. Even in late-S/G2 phases, NHEJ remained elevated relative to HR. Therefore, NHEJ is the major pathway for repairing endonuclease-induced DSBs in mammalian cells even in late-S/G2 phase, and does not compete with HR. | |||||
| 書誌情報 |
Environmental and Molecular Mutagenesis 巻 50, 号 9, p. 815-822, 発行日 2009-12 |
|||||
| ISSN | ||||||
| 収録物識別子タイプ | ISSN | |||||
| 収録物識別子 | 0893-6692 | |||||
