Mizuta R, LaSalle J M, Cheng H L, Shinohara A, Ogawa H, Copeland N, Jenkins N A, Lalande M, Alt F W
Howard Hughes Medical Institute, Children's Hospital, Boston, MA 02115, USA.
Proc Natl Acad Sci U S A. 1997 Jun 24;94(13):6927-32. doi: 10.1073/pnas.94.13.6927.
The human RAD51 protein is a homologue of the bacteria RecA and yeast RAD51 proteins that are involved in homologous recombination and DNA repair. RAD51 interacts with proteins involved in recombination and also with tumor suppressor proteins p53 and breast cancer susceptibility gene 1 (BRCA1). We have used the yeast two-hybrid system to clone murine cDNA sequences that encode two RAD51-associated molecules, RAB22 and RAB163. RAB163 encodes the C-terminal portion of mouse BRCA2, the homologue of the second breast cancer susceptibility gene protein in humans, demonstrating an in vitro association between RAD51 and BRCA2. RAB22 is a novel gene product that also interacts with RAD51 in vitro. To detect RAD51 interactions in vivo, we developed a transient nuclear focus assay that was used to demonstrate a complete colocalization of RAB22 with RAD51 in large nuclear foci.
人类RAD51蛋白是细菌RecA和酵母RAD51蛋白的同源物,它们参与同源重组和DNA修复。RAD51与参与重组的蛋白质相互作用,也与肿瘤抑制蛋白p53和乳腺癌易感基因1(BRCA1)相互作用。我们利用酵母双杂交系统克隆了编码两种RAD51相关分子RAB22和RAB163的小鼠cDNA序列。RAB163编码小鼠BRCA2的C末端部分,BRCA2是人类第二种乳腺癌易感基因蛋白的同源物,证明了RAD51与BRCA2之间的体外关联。RAB22是一种新型基因产物,它在体外也与RAD51相互作用。为了在体内检测RAD51的相互作用,我们开发了一种瞬时核灶分析方法,用于证明RAB22与RAD51在大的核灶中完全共定位。