Format

Send to

Choose Destination
Methods Mol Biol. 2016;1481:101-9. doi: 10.1007/978-1-4939-6393-5_11.

Reconstitution of the Cytoplasmic Regulation of the Wnt Signaling Pathway Using Xenopus Egg Extracts.

Author information

1
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.
2
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.

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.

KEYWORDS:

Axin; Egg extract; Proteasome; Ubiquitin; Xenopus laevis; β-Catenin destruction complex

PMID:
27590156
PMCID:
PMC5567996
DOI:
10.1007/978-1-4939-6393-5_11
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Springer Icon for PubMed Central
Loading ...
Support Center