Genome-wide identification of phospho-regulators of Wnt signaling in Drosophila

Development. 2015 Apr 15;142(8):1502-15. doi: 10.1242/dev.116715.

Abstract

Evolutionarily conserved intercellular signaling pathways regulate embryonic development and adult tissue homeostasis in metazoans. The precise control of the state and amplitude of signaling pathways is achieved in part through the kinase- and phosphatase-mediated reversible phosphorylation of proteins. In this study, we performed a genome-wide in vivo RNAi screen for kinases and phosphatases that regulate the Wnt pathway under physiological conditions in the Drosophila wing disc. Our analyses have identified 54 high-confidence kinases and phosphatases capable of modulating the Wnt pathway, including 22 novel regulators. These candidates were also assayed for a role in the Notch pathway, and numerous phospho-regulators were identified. Additionally, each regulator of the Wnt pathway was evaluated in the wing disc for its ability to affect the mechanistically similar Hedgehog pathway. We identified 29 dual regulators that have the same effect on the Wnt and Hedgehog pathways. As proof of principle, we established that Cdc37 and Gilgamesh/CK1γ inhibit and promote signaling, respectively, by functioning at analogous levels of these pathways in both Drosophila and mammalian cells. The Wnt and Hedgehog pathways function in tandem in multiple developmental contexts, and the identification of several shared phospho-regulators serve as potential nodes of control under conditions of aberrant signaling and disease.

Keywords: Hedgehog; In vivo RNAi screen; Notch; Wingless; Wnt.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Casein Kinase I / genetics
  • Casein Kinase I / metabolism
  • Cell Cycle Proteins / genetics
  • Cell Cycle Proteins / metabolism
  • Drosophila
  • Drosophila Proteins / genetics
  • Drosophila Proteins / metabolism*
  • Hedgehog Proteins / genetics
  • Hedgehog Proteins / metabolism
  • Molecular Chaperones / genetics
  • Molecular Chaperones / metabolism
  • Receptors, G-Protein-Coupled / genetics
  • Receptors, G-Protein-Coupled / metabolism
  • Receptors, Notch / genetics
  • Receptors, Notch / metabolism
  • Smoothened Receptor
  • Wings, Animal / metabolism
  • Wnt Proteins / genetics
  • Wnt Proteins / metabolism*
  • Wnt1 Protein / genetics
  • Wnt1 Protein / metabolism

Substances

  • Cdc37 protein, Drosophila
  • Cell Cycle Proteins
  • Drosophila Proteins
  • Hedgehog Proteins
  • Molecular Chaperones
  • N protein, Drosophila
  • Receptors, G-Protein-Coupled
  • Receptors, Notch
  • Smoothened Receptor
  • Wnt Proteins
  • Wnt1 Protein
  • smo protein, Drosophila
  • wg protein, Drosophila
  • hh protein, Drosophila
  • Casein Kinase I
  • gish protein, Drosophila