The role of BRCT domain in DNA damage response

BRCT结构域在DNA损伤反应中的作用

• 批准号: 18510045
• 负责人: KOBAYASHI Junya
• 金额: $ 2.59万
• 依托单位: Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 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-18510045/
• 关键词: Ionizing Radiation; Proteome; Cancer; Histone; DNA damage; Nucleolus; 遺伝子

Genome DNA suffers DNA damage by several stresses such as ionizing radiation and DV. Once DNA damage is generated into genome DNA, their cells detects this damage and induce growth arrest by cell cycle checkpoint system to repair damaged DNA, As the remain of DNA damage in genome DNA might lead to gene instability, tumorigenesis or apoptosis, it is much important to clarify the mechanism of DNA damage response. Recently, BRCT (BRCA1 C-Terminal) domain has been noticed in DNA damage response, bemuse some DNA damare response factors function by protein-protein interaction through their BRCT domain. However, the role of most BRCT domain proteins in DNA damage response is unknown Therefore, we identified interaction proteins with BRCT domain by proteomics analysis and investigated their the role of proteins in DNA damage response.NBS1, the responsible gene product for Nijmegen syndrome, interact with gamma-H2AX through FHA/BRCT domain, and this interaction is indispensable for focus format … More ion of NBS1/MRE11/RAD50 complex at DSB (DNA double-strand break) sites. Moreover, FHA/BRCT domain is also important for Homologous Recombination (HR) activity for DNA repair. TopBP1, which possesses BRCT domains, interacts with NBS1 dependently on the generation of DSB, and this interaction is essential to the DSB-dependent focus formation of TopBP1. Further, knockdown of TopBP1 reduced HR activity and sister chromatid exchange by HR, which suggests that TopBP1 function for HR. Furthermore, we identified nucleolin as a protein interacting with H2AX, which interact with BRCT domain of NBS1. Nucleolin interact with gamma-H2AX following generation of DNA damage. And, knockdown of nucleolin by siRNA increased gamma-H2AX and phosphorylation of ATM substrates and sensitivity to campthotecin (DNA damaging agent). Moreover, nucleolin knockdown cells showed decreases in HR activity. Thus, nuceloin might also be an important factor for HR. Therefore, the protein-protein interaction through BRCT domain could be much important for DNA damage response, particularly HR repair. As the mechanism of DNA damage response is suggested to be indispensable for barrier to tumorigenesis, we have to investigate more detailed role of BRCT domain in DNA damage response. Less

基因组DNA受到电离辐射和DV等多种胁迫的损伤。当基因组DNA发生损伤时，细胞会检测到损伤，并通过细胞周期检查点系统诱导细胞生长停滞，修复损伤的DNA。由于DNA损伤在基因组DNA中的残留可能导致基因不稳定、肿瘤发生或细胞凋亡，因此阐明DNA损伤反应的机制具有重要意义。近年来，BRCT（BRCA 1 C-Terminal）结构域在DNA损伤反应中的作用受到了广泛的关注，一些DNA损伤反应因子通过BRCT结构域发挥蛋白质-蛋白质相互作用。然而，大多数BRCT结构域蛋白在DNA损伤反应中的作用尚不清楚，因此，我们通过蛋白质组学分析鉴定了与BRCT结构域相互作用的蛋白质，并研究了它们在DNA损伤反应中的作用 ...更多信息 NBS 1/MRE 11/RAD 50复合物在DSB（DNA双链断裂）位点的离子。此外，FHA/BRCT结构域对于DNA修复的同源重组（HR）活性也是重要的。TopBP 1具有BRCT结构域，与NBS 1相互作用依赖于DSB的产生，并且这种相互作用对于TopBP 1的DSB依赖性焦点形成是必需的。此外，TopBP 1的敲低降低了HR活性和HR引起的姐妹染色单体交换，这表明TopBP 1对HR具有功能。此外，我们鉴定了核仁素作为一种与H2 AX相互作用的蛋白，它与NBS 1的BRCT结构域相互作用。核仁素与γ-H2 AX相互作用后产生DNA损伤。而且，通过siRNA敲低核仁素增加了γ-H2 AX和ATM底物的磷酸化以及对campthotecin（DNA损伤剂）的敏感性。此外，核仁素敲低细胞显示HR活性降低。因此，通过BRCT结构域的蛋白质-蛋白质相互作用可能在DNA损伤反应，特别是HR修复中起重要作用。由于DNA损伤反应的机制被认为是肿瘤发生的屏障所不可或缺的，因此我们需要更详细地研究BRCT结构域在DNA损伤反应中的作用。少

项目成果

TopBP1 associates with NBS1 and is involved in homologous recombination repair.

TopBP1 与 NBS1 结合并参与同源重组修复。

• 发表时间: 2007
• 期刊: Biochem Biophys Res Commun. 3;362(4)
• 作者: S. Sakamoto;K. Iijima;D. Mochizuki;K. Nakamura;K. Teshigawara;J. Kobayashi;S. Matsuura;H. Tauchi;K. Komatsu.;S. Sakamoto;K. Morishima
• 通讯作者: K. Morishima

Monoallelic BUBIB mutations and defective mitotic-spindle checkpoint in seven families with premature chromatid separation (PCS) syndrome

七个染色单体过早分离 (PCS) 综合征家系的单等位基因 BUBIB 突变和有丝分裂纺锤体检查点缺陷

• 发表时间: 2006
• 期刊: Am. J. Med. Genet. A 140
• 作者: Matsuura;S.;Matsumoto;Y.;Morishima;K.;Izumi;H.;Matsumoto;H.;Ito;E.;Tsutsui;Kobavashi;J.;Tauchi;H.;Kajiwara;Y.;Hama;S.;Kurisu;K.;Tahara;H.;Oshimura;M.;Komatsu;K.;Ikeuchi;T.;Kajii;T
• 通讯作者: T

Monoallelic BUB1B mutations and defective mitotic-spindle checkpoint in seven families with premature chromatid separation (PCS) syndrome

• DOI: 10.1002/ajmg.a.31069
• 发表时间: 2006-02-15
• 期刊: AMERICAN JOURNAL OF MEDICAL GENETICS PART A
• 影响因子: 2
• 作者: Matsuura, S;Matsumoto, Y;Kajii, T
• 通讯作者: Kajii, T

Functional interaction between histone H2AX and NBS1 on ATM-dependent DNA damage response.

组蛋白 H2AX 和 NBS1 在 ATM 依赖性 DNA 损伤反应中的功能相互作用。

• 发表时间: 2007
• 作者: 小林純也;他4名
• 通讯作者: 他4名

ATM is essential for DNA-flea phosphorylations at T2609 cluster upon DNA double strand break

ATM 对于 DNA 双链断裂后 T2609 簇上的 DNA-跳蚤磷酸化至关重要

• 发表时间: 2007
• 期刊: J Biol Chem 282
• 作者: Chen;BP.;Uematsu;N.;Kobavashi;J.;Lerenthal;Y.;Krempler;A.;Yajima;H.;Lobrich;M.;Shiloh;Y.;Chen;DJ
• 通讯作者: DJ