Smalley M J, Sara E, Paterson H, Naylor S, Cook D, Jayatilake H, Fryer L G, Hutchinson L, Fry M J, Dale T C
Developmental Biology, Section of Cell Biology and Experimental Pathology, Toby Robins Breakthrough Breast Cancer Research Centre, London, UK.
EMBO J. 1999 May 17;18(10):2823-35. doi: 10.1093/emboj/18.10.2823.
Axin promotes the phosphorylation of beta-catenin by GSK-3beta, leading to beta-catenin degradation. Wnt signals interfere with beta-catenin turnover, resulting in enhanced transcription of target genes through the increased formation of beta-catenin complexes containing TCF transcription factors. Little is known about how GSK-3beta-mediated beta-catenin turnover is regulated in response to Wnt signals. We have explored the relationship between Axin and Dvl-2, a member of the Dishevelled family of proteins that function upstream of GSK-3beta. Expression of Dvl-2 activated TCF-dependent transcription. This was blocked by co-expression of GSK-3beta or Axin. Expression of a 59 amino acid GSK-3beta-binding region from Axin strongly activated transcription in the absence of an upstream signal. Introduction of a point mutation into full-length Axin that prevented GSK-3beta binding also generated a transcriptional activator. When co-expressed, Axin and Dvl-2 co-localized within expressing cells. When Dvl-2 localization was altered using a C-terminal CAAX motif, Axin was also redistributed, suggesting a close association between the two proteins, a conclusion supported by co-immunoprecipitation data. Deletion analysis suggested that Dvl-association determinants within Axin were contained between residues 603 and 810. The association of Axin with Dvl-2 may be important in the transmission of Wnt signals from Dvl-2 to GSK-3beta.
轴蛋白(Axin)促进糖原合成酶激酶3β(GSK-3β)介导的β-连环蛋白磷酸化,导致β-连环蛋白降解。Wnt信号干扰β-连环蛋白的周转,通过增加含TCF转录因子的β-连环蛋白复合物的形成,导致靶基因转录增强。关于GSK-3β介导的β-连环蛋白周转如何响应Wnt信号进行调节,目前所知甚少。我们探究了Axin与Dvl-2之间的关系,Dvl-2是一种在GSK-3β上游起作用的散乱蛋白家族成员。Dvl-2的表达激活了TCF依赖的转录。这被GSK-3β或Axin的共表达所阻断。来自Axin的一个59个氨基酸的GSK-3β结合区域在没有上游信号的情况下强烈激活转录。将一个点突变引入全长Axin中,阻止GSK-3β结合,也产生了一种转录激活剂。当共表达时,Axin和Dvl-2在表达细胞内共定位。当使用C末端CAAX基序改变Dvl-2的定位时,Axin也重新分布,表明这两种蛋白之间存在紧密关联,这一结论得到了免疫共沉淀数据的支持。缺失分析表明,Axin内与Dvl结合的决定簇位于603和810位氨基酸残基之间。Axin与Dvl-2的结合在Wnt信号从Dvl-2传递到GSK-3β的过程中可能很重要。