Kishida S, Yamamoto H, Hino S, Ikeda S, Kishida M, Kikuchi A
Department of Biochemistry, Hiroshima University School of Medicine, Minami-ku, Hiroshima 734-8551, Japan.
Mol Cell Biol. 1999 Jun;19(6):4414-22. doi: 10.1128/MCB.19.6.4414.
The N-terminal region of Dvl-1 (a mammalian Dishevelled homolog) shares 37% identity with the C-terminal region of Axin, and this related region is named the DIX domain. The functions of the DIX domains of Dvl-1 and Axin were investigated. By yeast two-hybrid screening, the DIX domain of Dvl-1 was found to interact with Dvl-3, a second mammalian Dishevelled relative. The DIX domains of Dvl-1 and Dvl-3 directly bound one another. Furthermore, Dvl-1 formed a homo-oligomer. Axin also formed a homo-oligomer, and its DIX domain was necessary. The N-terminal region of Dvl-1, including its DIX domain, bound to Axin directly. Dvl-1 inhibited Axin-promoted glycogen synthase kinase 3beta-dependent phosphorylation of beta-catenin, and the DIX domain of Dvl-1 was required for this inhibitory activity. Expression of Dvl-1 in L cells induced the nuclear accumulation of beta-catenin, and deletion of the DIX domain abolished this activity. Although expression of Axin in SW480 cells caused the degradation of beta-catenin and reduced the cell growth rate, expression of an Axin mutant that lacks the DIX domain did not affect the level of beta-catenin or the growth rate. These results indicate that the DIX domains of Dvl-1 and Axin are important for protein-protein interactions and that they are necessary for the ability of Dvl-1 and Axin to regulate the stability of beta-catenin.
Dvl-1(一种哺乳动物中Dishevelled的同源物)的N端区域与Axin的C端区域有37%的同源性,这个相关区域被命名为DIX结构域。对Dvl-1和Axin的DIX结构域的功能进行了研究。通过酵母双杂交筛选,发现Dvl-1的DIX结构域与Dvl-3相互作用,Dvl-3是另一种哺乳动物Dishevelled相关蛋白。Dvl-1和Dvl-3的DIX结构域直接相互结合。此外,Dvl-1形成同源寡聚体。Axin也形成同源寡聚体,且其DIX结构域是必需的。Dvl-1的N端区域,包括其DIX结构域,直接与Axin结合。Dvl-1抑制Axin促进的糖原合酶激酶3β依赖的β-连环蛋白磷酸化,Dvl-1的DIX结构域对于这种抑制活性是必需的。在L细胞中表达Dvl-1诱导β-连环蛋白的核内积累,缺失DIX结构域则消除了这种活性。虽然在SW480细胞中表达Axin导致β-连环蛋白降解并降低细胞生长速率,但缺失DIX结构域的Axin突变体的表达并不影响β-连环蛋白水平或生长速率。这些结果表明,Dvl-1和Axin的DIX结构域对于蛋白质-蛋白质相互作用很重要,并且它们对于Dvl-1和Axin调节β-连环蛋白稳定性的能力是必需的。