title: Reconstitution of the Cytoplasmic Regulation of the Wnt Signaling Pathway Using Xenopus Egg Extracts.
abstract: 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.
authors: Department of Cell and Developmental Biology and Program in Developmental Biology, Vanderbilt University School of Medicine, PMB 407935 U-3218, Medical Research Building III, Nashville, TN, 37240, USA.
authors: Annastasia Simone
authors: AS
authors: Hyde
authors: Department of Cell and Developmental Biology and Program in Developmental Biology, Vanderbilt University School of Medicine, PMB 407935 U-3218, Medical Research Building III, Nashville, TN, 37240, USA.
authors: Brian I
authors: BI
authors: Hang
authors: Department of Cell and Developmental Biology and Program in Developmental Biology, Vanderbilt University School of Medicine, PMB 407935 U-3218, Medical Research Building III, Nashville, TN, 37240, USA. ethan.lee@vanderbilt.edu.
authors: Ethan
authors: E
authors: Lee
mesh_terms: D000818:Animals; D002453:Cell Cycle / Q000235:genetics; D042002:Chromatin Assembly and Disassembly / Q000235:genetics; D004261:DNA Replication / Q000235:genetics; D047108:Embryonic Development / Q000235:genetics; D008870:Microtubules / Q000235:genetics; D008967:Molecular Biology / Q000379:methods*; D009865:Oocytes / Q000378:metabolism; D059748:Proteolysis; D051153:Wnt Proteins; D060449:Wnt Signaling Pathway / Q000235:genetics; D014982:Xenopus laevis; D051176:beta Catenin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
chemical_list: D051153:Wnt Proteins; D051176:beta Catenin
keywords: Axin; Egg extract; Proteasome; Ubiquitin; Xenopus laevis; β-Catenin destruction complex