Kishida M, Koyama S, Kishida S, Matsubara K, Nakashima S, Higano K, Takada R, Takada S, Kikuchi A
Department of Biochemistry, Hiroshima University School of Medicine, Japan.
Oncogene. 1999 Jan 28;18(4):979-85. doi: 10.1038/sj.onc.1202388.
When Axin, a negative regulator of the Wnt signaling pathway, was expressed in COS cells, it coeluted with glycogen synthase kinase-3beta (GSK-3beta), beta-catenin, and adenomatous polyposis coli protein (APC) in a high molecular weight fraction on gel filtration column chromatography. In this fraction, GSK-3beta, beta-catenin, and APC were co-precipitated with Axin. Although beta-catenin was detected in the high molecular weight fraction in L cells on gel filtration column chromatography, addition of conditioned medium expressing Wnt-3a to the cells increased beta-catenin in the low molecular weight fraction. However, Wnt-3a-dependent accumulation of beta-catenin was greatly inhibited in L cells stably expressing Axin. Axin also suppressed Wnt-3a-dependent activation of Tcf-4 which binds to beta-catenin and acts as a transcription factor. These results suggest that Axin forms a complex with GSK-3beta, beta-catenin, and APC, resulting in the stimulation of the degradation of beta-catenin and that Wnt-3a induces the dissociation of beta-catenin from the Axin complex and accumulates beta-catenin.
当Wnt信号通路的负调节因子Axin在COS细胞中表达时,它在凝胶过滤柱色谱的高分子量组分中与糖原合酶激酶-3β(GSK-3β)、β-连环蛋白和腺瘤性息肉病大肠杆菌蛋白(APC)共同洗脱。在该组分中,GSK-3β、β-连环蛋白和APC与Axin共同沉淀。尽管在凝胶过滤柱色谱中L细胞的高分子量组分中检测到了β-连环蛋白,但向细胞中添加表达Wnt-3a的条件培养基会增加低分子量组分中的β-连环蛋白。然而,在稳定表达Axin的L细胞中,Wnt-3a依赖的β-连环蛋白积累受到极大抑制。Axin还抑制了与β-连环蛋白结合并作为转录因子起作用 的Tcf-4的Wnt-3a依赖性激活。这些结果表明,Axin与GSK-3β、β-连环蛋白和APC形成复合物,导致β-连环蛋白降解增加,并且Wnt-3a诱导β-连环蛋白从Axin复合物中解离并积累β-连环蛋白。