Bishop D K, Ear U, Bhattacharyya A, Calderone C, Beckett M, Weichselbaum R R, Shinohara A
Department of Radiation and Cellular Oncology, University of Chicago, Chicago, Illinois 60637, USA.
J Biol Chem. 1998 Aug 21;273(34):21482-8. doi: 10.1074/jbc.273.34.21482.
Rad51 is a member of a family of eukaryotic proteins related to the bacterial recombinational repair protein RecA. Rad51 protein localizes to multiple subnuclear foci in Chinese hamster ovary cells. Subnuclear Rad51 foci are induced by ionizing radiation or the DNA cross-linking agent cisplatin. Formation of these foci is likely to reflect assembly of a multimeric form of Rad51 that promotes DNA repair. Formation of damage-induced Rad51 foci does not occur in the Chinese hamster ovary cell line irs1SF, which is sensitive to DNA damaging agents. The Rad51 focus formation defect of irs1SF cells is corrected by a construct that encodes the repair protein Xrcc3. Xrcc3 is a human homolog of Rad51 previously isolated by virtue of its ability to correct the radiation sensitivity of irs1SF cells. Changes in the steady state level of Rad51 protein do not account for the irs1SF defect nor do they account for the appearance of foci following DNA damage. These results suggest that Xrcc3 is required for the assembly or stabilization of a multimeric form of Rad51 during DNA repair. Cell lines defective in two different components of DNA protein kinase formed Rad51 foci in response to damage, indicating DNA protein kinase is not required for damaged-induced mobilization of Rad51.
Rad51是真核生物蛋白质家族的一员,与细菌重组修复蛋白RecA相关。Rad51蛋白定位于中国仓鼠卵巢细胞的多个核内亚位点。核内亚位点的Rad51是由电离辐射或DNA交联剂顺铂诱导产生的。这些位点的形成可能反映了促进DNA修复的多聚体形式的Rad51的组装。在对DNA损伤剂敏感的中国仓鼠卵巢细胞系irs1SF中,不会出现由损伤诱导的Rad51位点的形成。编码修复蛋白Xrcc3的构建体可纠正irs1SF细胞的Rad51位点形成缺陷。Xrcc3是Rad51的人类同源物,先前因其能够纠正irs1SF细胞的辐射敏感性而被分离出来。Rad51蛋白稳态水平的变化既不能解释irs1SF的缺陷,也不能解释DNA损伤后位点的出现。这些结果表明,在DNA修复过程中,Xrcc3是Rad51多聚体形式组装或稳定所必需的。DNA蛋白激酶两个不同组分有缺陷的细胞系在受到损伤时会形成Rad51位点,这表明DNA蛋白激酶不是损伤诱导的Rad51动员所必需的。