Munemitsu S, Albert I, Rubinfeld B, Polakis P
Onyx Pharmaceuticals, Richmond, California 94806, USA.
Mol Cell Biol. 1996 Aug;16(8):4088-94. doi: 10.1128/MCB.16.8.4088.
Regulation of cell adhesion and cell signaling by beta-catenin occurs through a mechanism likely involving the targeted degradation of the protein. Deletional analysis was used to generate a beta-catenin refractory to rapid turnover and to examine its effects on complexes containing either cadherin or the adenomatous polyposis coli (APC) protein. The results show that amino-terminal deletion of beta-catenin results in a protein with increased stability that acts in a dominant fashion with respect to wild-type beta-catenin. Constitutive expression in AtT20 cells of a beta-catenin lacking 89 N-terminal amino acids (deltaN89beta-catenin) resulted in severely reduced levels of the more labile wild-type beta-catenin. The mutant beta-catenin was expressed at endogenous levels but displaced the vast majority of wild-type beta-catenin associated with N-cadherin. The deltaN89beta-catenin accumulated on the APC protein to a level 10-fold over that of wild-type beta-catenin and recruited a kinase into the APC complex. The kinase was highly active toward APC in vitro and promoted a sodium dodecyl sulfate gel band shift that was also evident for endogenous APC from cells expressing the mutant beta-catenin. Unlike wild-type beta-catenin, which partitions solely as part of a high-molecular-weight complex, the deltaN89 mutant protein also fractionated as a stable monomer, indicating that it had escaped the requirement to associate with other proteins. That similar N-terminal mutants of beta-catenin have been implicated in cellular transformation suggests that their abnormal association with APC may, in part, be responsible for this phenotype.
β-连环蛋白对细胞黏附和细胞信号传导的调节是通过一种可能涉及该蛋白质靶向降解的机制实现的。采用缺失分析来生成一种不易快速周转的β-连环蛋白,并研究其对包含钙黏蛋白或腺瘤性息肉病大肠杆菌(APC)蛋白的复合物的影响。结果表明,β-连环蛋白的氨基末端缺失会产生一种稳定性增加的蛋白质,该蛋白质相对于野生型β-连环蛋白以显性方式发挥作用。在AtT20细胞中组成型表达缺失89个氨基末端氨基酸的β-连环蛋白(δN89β-连环蛋白)导致更不稳定的野生型β-连环蛋白水平严重降低。突变型β-连环蛋白以内源性水平表达,但取代了与N-钙黏蛋白相关的绝大多数野生型β-连环蛋白。δN89β-连环蛋白在APC蛋白上的积累水平比野生型β-连环蛋白高10倍,并将一种激酶招募到APC复合物中。该激酶在体外对APC具有高度活性,并促进了十二烷基硫酸钠凝胶条带迁移,这在表达突变型β-连环蛋白的细胞的内源性APC中也很明显。与仅作为高分子量复合物一部分进行分配的野生型β-连环蛋白不同,δN89突变蛋白也以稳定单体的形式分级分离,这表明它不再需要与其他蛋白质结合。β-连环蛋白的类似氨基末端突变体与细胞转化有关,这表明它们与APC的异常结合可能在一定程度上导致了这种表型。