Skip to main content

Protein–Protein Interaction Techniques: Dissect PCP Signaling in Xenopus

  • Protocol
  • First Online:
Planar Cell Polarity

Part of the book series: Methods in Molecular Biology ((MIMB,volume 839))

Abstract

The planar cell polarity (PCP) pathway is a β-catenin-independent branch of the Wnt signaling cascade. In vertebrate embryos PCP signaling regulates morphogenetic events including convergent extension (CE) movements during gastrualtion. Xenopus embryo has been established as an excellent model system to dissect PCP signaling in vertebrates because morphogenetic cell behaviors including CE can easily be monitored in vivo. Xenopus Paraxial protocadherin (xPAPC) is a transmembrane protein which serves as a link between patterning factors in the Spemann’s organizer and regulators of the morphogenetic movements. xPAPC regulates morphogenesis in part by modulating cell adhesion and PCP signaling. Here two methods, GST pull-down assay and yeast two-hybrid assay, are described for the identification of xPAPC interacting proteins to elucidate the mechanism by which xPAPC regulates PCP signaling.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Zallen JA. (2007) Planar polarity and tissue morphogenesis. Cell 129:1051–1063

    Article  PubMed  CAS  Google Scholar 

  2. Jones C, Chen P. (2008) Primary cilia in planar cell polarity regulation of the inner ear. Curr Top Dev Biol 85:197–224

    Article  PubMed  CAS  Google Scholar 

  3. Simons M, Mlodzik M. (2008) Planar cell polarity signaling: from fly development to human disease. Annu Rev Genet 42:517–540

    Article  PubMed  CAS  Google Scholar 

  4. Wang Y. (2009) Wnt/Planar cell polarity signaling: a new paradigm for cancer therapy. Mol Cancer Ther 8:2103–2109

    Article  PubMed  CAS  Google Scholar 

  5. Wang Y, Steinbeisser H. (2009) Molecular basis of morphogenesis during vertebrate gastrulation. Cell Mol Life Sci 66:2263–2273

    Article  PubMed  CAS  Google Scholar 

  6. Medina A, Swain RK, Kuerner KM, Steinbeisser H. (2004) Xenopus paraxial protocadherin has signaling functions and is involved in tissue separation. EMBO J 23:3249–3258

    Article  PubMed  CAS  Google Scholar 

  7. Unterseher F, Hefele, JA, Giehl K, De Robertis EM, Wedlich D, Schambony A. (2004) Paraxial protocadherin coordinates cell polarity during convergent extension via Rho A and JNK. EMBO J 23:3259–3269

    Article  PubMed  CAS  Google Scholar 

  8. Wang Y, Janicki P, Köster I, Berger CD, Wenzl C, Grosshans J, Steinbeisser H. (2008) Xenopus Paraxial Protocadherin regulates morphogenesis by antagonizing Sprouty. Genes Dev 22:878–883

    Article  PubMed  CAS  Google Scholar 

  9. Debonneville C, Staub O. (2004) Participation of the ubiquitin-conjugating enzyme UBE2E3 in Nedd4-2-dependent regulation of the epithelial Na  +  channel. Mol Cell Biol 24:2397–2409

    Article  PubMed  CAS  Google Scholar 

  10. Iouzalen N, Camonis J, Moreau J. (1998) Identification and Characterization in Xenopus of XsmgGDS, a RalB Binding Protein. Biochemical and Biophysical Research Commu­nications 250:359–363

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

I am deeply indebted to Herbert Steinbeisser for helpful discussion and critical reading of the manuscript. I thank O. Staub for kindly providing yeast two-hybrid library and J. Moreau for pNLX3 plasmid.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Yingqun Wang .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2012 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Wang, Y. (2012). Protein–Protein Interaction Techniques: Dissect PCP Signaling in Xenopus . In: Turksen, K. (eds) Planar Cell Polarity. Methods in Molecular Biology, vol 839. Springer, New York, NY. https://doi.org/10.1007/978-1-61779-510-7_3

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-510-7_3

  • Published:

  • Publisher Name: Springer, New York, NY

  • Print ISBN: 978-1-61779-509-1

  • Online ISBN: 978-1-61779-510-7

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics