Pitchfork and Gprasp2 Target Smoothened to the Primary Cilium for Hedgehog Pathway Activation

Bomi Jung; Daniela Padula; Ingo Burtscher; Cedric Landerer; Dominik Lutter; Fabian Theis; Ana C Messias; Arie Geerlof; Michael Sattler; Elisabeth Kremmer; Karsten Boldt; Marius Ueffing; Heiko Lickert

doi:10.1371/journal.pone.0149477

Pitchfork and Gprasp2 Target Smoothened to the Primary Cilium for Hedgehog Pathway Activation

PLoS One. 2016 Feb 22;11(2):e0149477. doi: 10.1371/journal.pone.0149477. eCollection 2016.

Authors

Bomi Jung^{1

2

3}, Daniela Padula^{1

2

3}, Ingo Burtscher^{1

2

3}, Cedric Landerer^{4

5}, Dominik Lutter^{4

6}, Fabian Theis⁴, Ana C Messias^{7

8}, Arie Geerlof⁷, Michael Sattler^{7

8}, Elisabeth Kremmer⁹, Karsten Boldt^{10

11}, Marius Ueffing^{10

11}, Heiko Lickert^{1

2

3}

Affiliations

¹ Institute of Diabetes and Regeneration Research, Helmholtz Zentrum München, München-Neuherberg, Germany.
² Institute of Stem Cell Research, Helmholtz Zentrum München, München-Neuherberg, Germany.
³ German Center for Diabetes Research (DZD), 85764 München-Neuherberg, Germany.
⁴ Institute of Computational Biology, Helmholtz Zentrum München, München-Neuherberg, Germany.
⁵ Department of Ecology & Evolutionary Biology, University of Tennessee, Knoxville TN 37996, United States of America.
⁶ Institute of Diabetes and Adipositas, Helmholtz Zentrum München, München-Neuherberg, Germany.
⁷ Institute of Structural Biology, Helmholtz Zentrum München, München-Neuherberg, Germany.
⁸ Center for Integrated Protein Science Munich at Biomolecular NMR Spectroscopy, Department Chemie, Technische Universität München, 85747 Garching, Germany.
⁹ Institute of Molecular Immunology, Helmholtz Zentrum München, München-Neuherberg, Germany.
¹⁰ Department of Protein Science, Helmholtz Zentrum München, München-Neuherberg, Germany.
¹¹ Centre of Ophthalmology, Institute for Ophthalmology Research, University of Tuebingen, 72076 Tuebingen, Germany.

Abstract

The seven-transmembrane receptor Smoothened (Smo) activates all Hedgehog (Hh) signaling by translocation into the primary cilia (PC), but how this is regulated is not well understood. Here we show that Pitchfork (Pifo) and the G protein-coupled receptor associated sorting protein 2 (Gprasp2) are essential components of an Hh induced ciliary targeting complex able to regulate Smo translocation to the PC. Depletion of Pifo or Gprasp2 leads to failure of Smo translocation to the PC and lack of Hh target gene activation. Together, our results identify a novel protein complex that is regulated by Hh signaling and required for Smo ciliary trafficking and Hh pathway activation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Line
Cilia / genetics
Cilia / metabolism
Hedgehog Proteins / genetics
Hedgehog Proteins / metabolism*
Homeodomain Proteins / genetics
Homeodomain Proteins / metabolism*
Intracellular Signaling Peptides and Proteins
Mice
Protein Transport / physiology
Receptors, G-Protein-Coupled / genetics
Receptors, G-Protein-Coupled / metabolism*
Signal Transduction / physiology*
Smoothened Receptor

Substances

Gprasp2 protein, mouse
Hedgehog Proteins
Homeodomain Proteins
Intracellular Signaling Peptides and Proteins
Receptors, G-Protein-Coupled
Smo protein, mouse
Smoothened Receptor
pitchfork protein, mouse

Grants and funding

This work was funded in part by the German Center for Diabetes Research (DZD e.V.) and the Helmholtz Alliance ICEMED - Imaging and Curing Environmental Metabolic Diseases. Financial support was additionally provided by an Emmy-Noether fellowship from the German Research Foundation and an ERC starting grant awarded to HL.