Analysis of Casein kinase I function in the Wnt signal-mediated regulation of Armadillo family protein degradation.

酪蛋白激酶 I 在 Wnt 信号介导的犰狳家族蛋白降解调节中的功能分析。

• 批准号: 15570113
• 负责人: YANAGAWA Shin-ichi
• 金额: $ 2.37万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15570113/
• 关键词: Wnt; Protein Degradation; β-catenin; Casein kinase I; Signal Transduction; Casein Kinase I; Wn^+; Wingless; casein kinase I; GSK-3B; Armadillo; RNAi; Signal Transduction

The Wnt family of secretory protein plays pivotal roles in a number of basic developmental processes. In addition, numerous mutations in components of the Wnt pathway, such as apc, axin, β-catenin, are oncogenic. It is well established that regulation of Arm protein levels through ubiquitin-mediated degradation plays a central role in the Wingless (Wg) signaling. Although Zeste White 3(Zw3)-mediated Arm phosphorylation has been implicated in its degradation, we have recently shown that Casein kinase Ia(CKIα) also phosphorylates Arm and induces its degradation. Thus, CKIα is functioning as a very strong negative regulator of the canonical Wnt/Wg signaling pathway.But it remains unclear how CKIα and Zw3 as well as other components of the Arm degradation complex regulate Arm phosphorylation in response to Wg. In particular, whether Wg signaling suppresses CKIα- or Zw3-mediated Arm phosphorylaytion in vivo is unknown. To clarify these issues, we performed a series of RNA interference(RNAi)-based analyses in Drosophila S2R+ cells using antibodies that specifically recognize Arm phosphorylated at different serine residues. These analyses revealed that Arm phosphorylation at serine56, and at threonine52, serine48, and serine44, is mediated by CKIα and Zw3, respectively and that Zw3-directed Arm phosphorylation requires CKIα-mediated priming phosphorylation. Daxin stimulates Zw3- but not CKIα-mediated Arm phosphorylation. Wg suppresses Zw3- but not CKIα-mediated Arm phosphorylation, indicating that a vital regulatory step in Wg signaling is Zw3-mediated Arm phosphorylation. In addition, further RNAi-based analyses of the other aspects of the Wg pathway clarified that Wg-induced Dishevelled phosphoylation is due to CKIα and that Presenilin and Protein kinaseA play little part in the regulation of Arm protein levels in Drosophila tissue culture cells.

分泌蛋白Wnt家族在许多基本发育过程中起着关键作用。此外，Wnt通路的组分中的许多突变，例如apc、轴蛋白、β-连环蛋白，是致癌的。已经确定，通过泛素介导的降解调节Arm蛋白水平在无翼（Wg）信号传导中起核心作用。尽管Zeste白色3（Zw 3）介导的Arm磷酸化与其降解有关，但我们最近发现酪蛋白激酶Ia（CKIα）也使Arm磷酸化并诱导其降解。因此，CKIα在经典Wnt/Wg信号通路中起着非常强的负调节作用，但目前尚不清楚CKIα和Zw 3以及Arm降解复合物的其他组分如何调节Arm磷酸化以响应Wg。特别是，Wg信号传导是否抑制CKIα或Zw 3介导的Arm体内磷酸化尚不清楚。为了澄清这些问题，我们使用特异性识别在不同丝氨酸残基磷酸化的Arm的抗体在果蝇S2 R+细胞中进行了一系列基于RNA干扰（RNAi）的分析。这些分析表明，Arm丝氨酸56、苏氨酸52、丝氨酸48和丝氨酸44的磷酸化分别由CKIα和Zw 3介导，Zw 3介导的Arm磷酸化需要CKIα介导的引发磷酸化。Daxin刺激Zw 3-而不是CKIα-介导的Arm磷酸化。Wg抑制Zw 3介导的Arm磷酸化，但不抑制CKIα介导的Arm磷酸化，表明Wg信号传导中的重要调节步骤是Zw 3介导的Arm磷酸化。此外，对Wg途径其他方面的进一步基于RNAi的分析阐明，Wg诱导的Dishevelled磷酸化是由于CKIα，而早老素和蛋白激酶A在果蝇组织培养细胞中调节Arm蛋白水平方面几乎没有作用。

项目成果

Axin and the Axin/Arrow-binding protein DCAP mediate glucose-glycogen metabolism.

Axin 和 Axin/Arrow 结合蛋白 DCAP 介导葡萄糖-糖原代谢。

• 发表时间: 2003
• 期刊: Biochemical and Biophysical Research Communications 304
• 作者: Hiroto Yamazaki;Shin-ichi Yanagawa
• 通讯作者: Shin-ichi Yanagawa

Hiroko Matsubayashi: "Biochemical characterization of the Drosophila Wingless Signaling pathway Based on RNA Interference"Molecular and Cellular Biology. 24巻・5号. 2012-2024 (2004)

Hiroko Matsubayashi：“基于 RNA 干扰的果蝇 Wingless 信号通路的生化表征”《分子与细胞生物学》第 24 卷，第 5 期。2012-2024 年（2004 年）

Hiroto Yamazaki: "Axin and the Axin/Arrow-binding protein DCAP mediate glucose-glycogen metabolism"Biochemical and Biophysical Research Communications. 304巻. 229-235 (2003)

Hiroto Yamazaki：“Axin 和 Axin/Arrow 结合蛋白 DCAP 介导葡萄糖-糖原代谢”，生物化学和生物物理研究通讯，第 304 卷，229-235（2003 年）。

Biochemical characterization of the Drosophila Wingless Pathway based on RNA interference.

基于 RNA 干扰的果蝇无翅途径的生化表征。

• 发表时间: 2004
• 期刊: Molecular and Cellular Biology 24
• 作者: Hiroko Matsubayashi;Sonoka Sese;Jong-Seo Lee;Tadaoki Shirakawa;Takeshi Iwatsubo;Taisuke Tomita;Shin-ichi Yanagawa
• 通讯作者: Shin-ichi Yanagawa

Axin and the Axin/Arrow-binding protein DCAP mediate glucose-glycogen metabolism

Axin 和 Axin/Arrow 结合蛋白 DCAP 介导葡萄糖-糖原代谢

• 发表时间: 2003
• 期刊: Biochemical and Biophysical Research Communications 304
• 作者: Hiroto Yamazaki;Shin-ichi Yanagawa
• 通讯作者: Shin-ichi Yanagawa