Yuan Shyng-Shiou F, Chang Hsueh-Ling, Lee Eva Y-H P
Department of Obstetrics and Gynecology, Kaohsiung Medical University Hospital, Kaohsiung 807, Taiwan, ROC.
Mutat Res. 2003 Apr 9;525(1-2):85-92. doi: 10.1016/s0027-5107(03)00009-5.
In eukaryotes, DNA double-strand breaks (DSBs) can be repaired by either non-homologous end-joining (NHEJ) or homologous recombination (HR) pathways. Rad50 protein is a component of the Rad50/NBS1/Mre11 nuclease complex that functions in both the NHEJ and recombinational repair of DNA DSBs. On the other hand, Rad51 protein, a homolog of bacterial RecA and a member of the Rad52 epistasis group, plays a crucial role exclusively in the recombinational repair pathway. We analyzed the effects of cell cycle progression and genetic background on the ionizing radiation (IR)-induced Rad51 and Rad50 repair focus formation. Herein, we demonstrated that IR-induced Rad51, but not Rad50, nuclear focus formation was cell cycle-dependent. Furthermore, IR-induced Rad51 focus formation was defective in AT and c-Abl(-/-) cells, but not wild type or NBS cells. A decreased and delayed formation of Rad51 foci-containing nuclei was observed in AT cells upon IR, whereas in c-Abl(-/-) cells a decreased but not delayed formation of Rad51 foci-containing nuclei was observed. In conclusion, effective and prompt IR-induced Rad51 focus formation is cell cycle-regulated and requires both ATM and c-Abl.
在真核生物中,DNA双链断裂(DSB)可通过非同源末端连接(NHEJ)或同源重组(HR)途径进行修复。Rad50蛋白是Rad50/NBS1/Mre11核酸酶复合物的一个组成部分,该复合物在DNA DSB的NHEJ和重组修复中均发挥作用。另一方面,Rad51蛋白是细菌RecA的同源物,属于Rad52上位性组的成员,仅在重组修复途径中起关键作用。我们分析了细胞周期进程和遗传背景对电离辐射(IR)诱导的Rad51和Rad50修复灶形成的影响。在此,我们证明了IR诱导的Rad51而非Rad50核灶形成是细胞周期依赖性的。此外,IR诱导的Rad51灶形成在共济失调毛细血管扩张症(AT)细胞和c-Abl基因敲除(-/-)细胞中存在缺陷,但在野生型或NBS细胞中则没有。IR照射后,在AT细胞中观察到含Rad51灶的细胞核形成减少且延迟,而在c-Abl(-/-)细胞中观察到含Rad51灶的细胞核形成减少但没有延迟。总之,IR诱导的有效且迅速的Rad51灶形成受细胞周期调控,并且需要ATM和c-Abl两者。