論文 - 詳細
| RRC ID | 18917 |
|---|---|
| 著者 | Kamdar RP, Matsumoto Y. |
| タイトル | Radiation-induced XRCC4 association with chromatin DNA analyzed by biochemical fractionation. |
| ジャーナル | J Radiat Res |
| Abstract |
XRCC4, in association with DNA ligase IV, is thought to play a critical role in the ligation of two DNA ends in DNA double-strand break (DSB) repair through non-homologous end-joining (NHEJ) pathway. In the present study, we captured radiation-induced chromatin-recruitment of XRCC4 by biochemical fractionation using detergent Nonidet P-40. A subpopulation of XRCC4 changed into a form that is resistant to the extraction with 0.5% Nonidet P-40-containing buffer after irradiation. This form of XRCC4 was liberated by micrococcal nuclease treatment, indicating that it had been tethered to chromatin DNA. This chromatin-recruitment of XRCC4 could be seen immediately (< 0.1 hr) after irradiation and remained up to 4 hr after 20 Gy irradiation. It was seen even after irradiation of small doses, i.e., 2 Gy, but the residence of XRCC4 on chromatin was very transient after 2 Gy irradiation, returning to near normal level in 0.2-0.5 hr after irradiation. The chromatin-bound XRCC4 represented only approximately 1% of total XRCC4 molecules even after 20 Gy irradiation and the quantitative analysis using purified protein as the reference suggested that only a few XRCC4-DNA ligase IV complexes were recruited to each DNA end. We further show that the chromatin-recruitment of XRCC4 was not attenuated by wortmannin, an inhibitor of DNA-PK, or siRNA-mediated knockdown of the DNA-PK catalytic subunit (DNA-PKcs), indicating that this process does not require DNA-PKcs. These results would provide us with useful experimental tools and important insights to understand the DNA repair process through NHEJ pathway. |
| 巻・号 | 51(3) |
| ページ | 303-13 |
| 公開日 | 2010-1-1 |
| DOI | 10.1269/jrr.09146 |
| PII | JST.JSTAGE/jrr/09146 |
| PMID | 20448413 |
| MeSH | Androstadienes / pharmacology Animals Catalytic Domain Cell Line, Tumor Cell Proliferation Chromatin / chemistry Chromatin / genetics* Chromatin / metabolism DNA / genetics* DNA / metabolism DNA-Activated Protein Kinase / metabolism DNA-Binding Proteins / metabolism* Dose Fractionation, Radiation* Dose-Response Relationship, Radiation Mice Micrococcal Nuclease / metabolism RNA, Small Interfering / metabolism Wortmannin X-Rays |
| IF | 1.95 |
| 引用数 | 15 |
| WOS 分野 | BIOLOGY RADIOLOGY, NUCLEAR MEDICINE & MEDICAL IMAGING |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 各媒体での言及数の合計 | 0 |
| リソース情報 | |
| ヒト・動物細胞 | M1(RCB0169) |