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Wnt Signaling pp 101-109 | Cite as

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

  • Annastasia Simone Hyde
  • Brian I. Hang
  • Ethan LeeEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1481)

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.

Key words

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

Notes

Acknowledgements

We thank Laurie Lee for critical reading of the manuscript. ASH is supported by a fellowship from the NIH Molecular Endocrinology Training Program. EL is supported by the NIH grants R01GM081635 and R01GM103926.

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Annastasia Simone Hyde
    • 1
  • Brian I. Hang
    • 1
  • Ethan Lee
    • 1
    Email author
  1. 1.Department of Cell and Developmental Biology and Program in Developmental BiologyVanderbilt University School of MedicineNashvilleUSA

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