Takaku Motoki, Machida Shinichi, Hosoya Noriko, Nakayama Shugo, Takizawa Yoshimasa, Sakane Isao, Shibata Takehiko, Miyagawa Kiyoshi, Kurumizaka Hitoshi
Laboratory of Structural Biology, Graduate School of Advanced Science and Engineering, and Consolidated Research Institute for Advanced Science and Medical Care, Waseda University, 2-2 Wakamatsu-cho, Shinjuku-ku, Tokyo 162-8480, Japan.
J Biol Chem. 2009 May 22;284(21):14326-36. doi: 10.1074/jbc.M807715200. Epub 2009 Mar 26.
The RAD51 protein is a central player in homologous recombinational repair. The RAD51B protein is one of five RAD51 paralogs that function in the homologous recombinational repair pathway in higher eukaryotes. In the present study, we found that the human EVL (Ena/Vasp-like) protein, which is suggested to be involved in actin-remodeling processes, unexpectedly binds to the RAD51 and RAD51B proteins and stimulates the RAD51-mediated homologous pairing and strand exchange. The EVL knockdown cells impaired RAD51 assembly onto damaged DNA after ionizing radiation or mitomycin C treatment. The EVL protein alone promotes single-stranded DNA annealing, and the recombination activities of the EVL protein are further enhanced by the RAD51B protein. The expression of the EVL protein is not ubiquitous, but it is significantly expressed in breast cancer-derived MCF7 cells. These results suggest that the EVL protein is a novel recombination factor that may be required for repairing specific DNA lesions, and that may cause tumor malignancy by its inappropriate expression.
RAD51蛋白是同源重组修复过程中的核心参与者。RAD51B蛋白是高等真核生物同源重组修复途径中发挥作用的五个RAD51旁系同源物之一。在本研究中,我们发现人类EVL(Ena/Vasp样)蛋白,据推测其参与肌动蛋白重塑过程,却意外地与RAD51和RAD51B蛋白结合,并刺激RAD51介导的同源配对和链交换。EVL基因敲低的细胞在受到电离辐射或丝裂霉素C处理后,RAD51组装到受损DNA上的过程受损。单独的EVL蛋白可促进单链DNA退火,并且RAD51B蛋白可进一步增强EVL蛋白的重组活性。EVL蛋白的表达并非普遍存在,而是在乳腺癌来源的MCF7细胞中显著表达。这些结果表明,EVL蛋白是一种新型重组因子,可能是修复特定DNA损伤所必需的,并且可能因其表达不当而导致肿瘤恶性化。