Characterization of mouse dishevelled (Dvl) proteins in Wnt/wingless signaling pathway
Characterization of mouse dishevelled (Dvl) proteins in Wnt/wingless signaling pathway
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DOI：
10.1074/jbc.274.30.21464
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发表时间：
1999-07-23
影响因子：
4.8
通讯作者：
Yanagawa, S
中科院分区：
文献类型：
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作者：
Lee, JS;Ishimoto, A;Yanagawa, S
Dheveled(Dsh)基因家族编码与Wnt/Wingless(Wg)信号相关的细胞质蛋白。为了证明Dsh家族蛋白的功能保守性，在小鼠和果蝇细胞培养系统中对果蝇Dsh的两个小鼠同源物DVL-1和DVL-2进行了生化鉴定。我们发现，在小鼠NIH3T3、L和C57 MG细胞中，可溶性WNT-3A处理导致DVL蛋白过度磷酸化，并伴随细胞质β-连环素水平的升高。这与我们在果蝇翼盘细胞系克隆中的发现很好地一致；Wg处理诱导了]Dsh(Yanagawa，S.，van Leeuwen，F.，Wodarz，A.，Klingensmith，J.和Nusse，R.(1995)基因Dev的过度磷酸化。9、1087-1097)。此外，我们发现小鼠DVL蛋白像Dsh一样影响果蝇Wg信号的下游成分；在Clone-8细胞中过表达DVL蛋白会导致Armadillo(果蝇β-catenin同源物)和果蝇E-cadherin水平升高，DVL蛋白本身过度磷酸化，并抑制Zust-White 3激酶介导的微管结合蛋白Tan的磷酸化。此外，酪蛋白激酶II与DVL蛋白共沉淀，DVL蛋白在这些免疫复合物中被磷酸化。这些结果直接证明了Dsh家族蛋白在Wnt/Wg信号通路中介导了一系列保守的生化过程。
The dishevelled (dsh) gene family encodes cytoplasmic proteins that have been implicated in Wnt/Wingless (Wg) signaling. To demonstrate functional conservation of Dsh family proteins, two mouse homologs of Drosophila Dsh, Dvl-1 and Dvl-2, were biochemically characterized in mouse and Drosophila cell culture systems. We found that treatment with a soluble Wnt-3A leads to hyperphosphorylation of Dvl proteins and a concomitant elevation of the cytoplasmic beta-catenin levels in mouse NIH3T3, L, and C57MG cells. This coincides well with our finding in a Drosophila wing disc cell line, clones, that; Wg treatment induced hyperphosphorylation of]Dsh (Yanagawa, S., van Leeuwen, F., Wodarz, A., Klingensmith, J., and Nusse, R. (1995) Genes Dev. 9, 1087-1097). Furthermore, we showed that mouse Dvl proteins affect downstream components of Drosophila Wg signaling as Dsh does; overexpression of Dvl proteins in clone-8 cells results in elevation of Armadillo (Drosophila homolog of beta-catenin) and Drosophila E-cadherin levels, hyperphosphorylation of Dvl proteins themselves, and inhibition of Zeste-White3 kinase-mediated phosphorylation of a microtubule-binding protein, Tan. In addition, casein kinase II was shown to coimmunoprecipitate with Dvl proteins, and Dvl proteins were phosphorylated in these immune complexes. These results are direct evidence that Dsh family proteins mediate a set of conserved biochemical processes in the Wnt/Wg signaling pathway.