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.
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References
Zallen JA. (2007) Planar polarity and tissue morphogenesis. Cell 129:1051–1063
Jones C, Chen P. (2008) Primary cilia in planar cell polarity regulation of the inner ear. Curr Top Dev Biol 85:197–224
Simons M, Mlodzik M. (2008) Planar cell polarity signaling: from fly development to human disease. Annu Rev Genet 42:517–540
Wang Y. (2009) Wnt/Planar cell polarity signaling: a new paradigm for cancer therapy. Mol Cancer Ther 8:2103–2109
Wang Y, Steinbeisser H. (2009) Molecular basis of morphogenesis during vertebrate gastrulation. Cell Mol Life Sci 66:2263–2273
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
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
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
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
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
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.
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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
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DOI: https://doi.org/10.1007/978-1-61779-510-7_3
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