Reconstitution of the Cytoplasmic Regulation of the Wnt Signaling Pathway Using Xenopus Egg Extracts.
Reconstitution of the Cytoplasmic Regulation of the Wnt Signaling Pathway Using Xenopus Egg Extracts.
复制标题
使用异爪蟾卵提取物对Wnt信号通路的细胞质调节重构。
DOI：
10.1007/978-1-4939-6393-5_11
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
Lee E
中科院分区：
文献类型：
--
作者：
Hyde AS;Hang BI;Lee E
β-连环蛋白周转的调节是控制 Wnt 信号通路激活的核心机制。该途径的所有成分都存在于非洲爪蟾的早期胚胎中，非洲爪蟾卵提取物已被用于重现复杂的生物反应，例如微管动力学、DNA 复制、染色质组装和细胞周期的阶段。在此，我们描述了一种使用爪蟾卵提取物中的放射性标记和荧光素酶融合蛋白分析 β-连环蛋白降解的生化方法。我们发现，在这样的生化系统中，细胞质 β-catenin 降解受到 Wnt 途径的可溶性成分以及小分子的调节。
The regulation of β-catenin turnover is the central mechanism governing activation of the Wnt signaling pathway. All components of the pathway are present in the early embryo of Xenopus laevis, and Xenopus egg extracts have been used to recapitulate complex biological reactions such as microtubule dynamics, DNA replication, chromatin assembly, and phases of the cell cycle. Herein, we describe a biochemical method for analyzing β-catenin degradation using radiolabeled and luciferase-fusion proteins in Xenopus egg extracts. We show that in such a biochemical system, cytoplasmic β-catenin degradation is regulated by soluble components of the Wnt pathway as well as small molecules.