Miki T, Smith C L, Long J E, Eva A, Fleming T P
Laboratory of Cellular and Molecular Biology, National Cancer Institute, Bethesda, Maryland 20892.
Nature. 1993 Apr 1;362(6419):462-5. doi: 10.1038/362462a0.
We have developed an efficient expression cloning system that allows rapid isolation of complementary DNAs able to induce the transformed phenotype. We searched for molecules expressed in epithelial cells and possessing transforming potential to fibroblasts, and cloned a cDNA for the normal receptor of a growth factor secreted by NIH/3T3 cells. Here we report a second novel transforming gene, ect2. The isolated cDNA is activated by amino-terminal truncation of the normal product. The Ect2 protein has sequence similarity within a central core of 255 amino acids with the products of the breakpoint cluster gene, bcr (ref. 5), the yeast cell cycle gene, CDC24 (ref. 6), and the dbl oncogene. Each of these genes encodes regulatory molecules or effectors for Rho-like small GTP-binding proteins. The baculovirus-expressed Ect2 protein could bind highly specifically to Rho and Rac proteins, whereas the dbl product showed broader binding specificity to Rho family proteins. Thus ect2 is a new member of an expanding family, whose products have transforming properties and interact with Rho-like proteins of the Ras superfamily.
我们开发了一种高效的表达克隆系统,该系统能够快速分离出可诱导转化表型的互补DNA。我们寻找在上皮细胞中表达且对成纤维细胞具有转化潜力的分子,并克隆了NIH/3T3细胞分泌的一种生长因子的正常受体的cDNA。在此,我们报告第二个新型转化基因ect2。分离出的cDNA因正常产物的氨基末端截短而被激活。Ect2蛋白在255个氨基酸的中央核心区域与断裂点簇基因bcr(参考文献5)、酵母细胞周期基因CDC24(参考文献6)以及dbl癌基因的产物具有序列相似性。这些基因中的每一个都编码Rho样小GTP结合蛋白的调节分子或效应器。杆状病毒表达的Ect2蛋白能高度特异性地结合Rho和Rac蛋白,而dbl产物对Rho家族蛋白表现出更广泛的结合特异性。因此,ect2是一个不断扩大的家族的新成员,该家族的产物具有转化特性并与Ras超家族的Rho样蛋白相互作用。