Time-resolved proteomics profiling of the ciliary Hedgehog response.

Abstract

The primary cilium is a signaling compartment that interprets Hedgehog signals through changes of its protein, lipid, and second messenger compositions. Here, we combine proximity labeling of cilia with quantitative mass spectrometry to unbiasedly profile the time-dependent alterations of the ciliary proteome in response to Hedgehog. This approach correctly identifies the three factors known to undergo Hedgehog-regulated ciliary redistribution and reveals two such additional proteins. First, we find that a regulatory subunit of the cAMP-dependent protein kinase (PKA) rapidly exits cilia together with the G protein-coupled receptor GPR161 in response to Hedgehog, and we propose that the GPR161/PKA module senses and amplifies cAMP signals to modulate ciliary PKA activity. Second, we identify the phosphatase Paladin as a cell type-specific regulator of Hedgehog signaling that enters primary cilia upon pathway activation. The broad applicability of quantitative ciliary proteome profiling promises a rapid characterization of ciliopathies and their underlying signaling malfunctions.

• Authors: Elena A May, Marian Kalocsay, Inès Galtier D'Auriac, Patrick S Schuster, Steven P Gygi, Maxence V Nachury, David U Mick
• Journal: The Journal of cell biology (J Cell Biol) Vol. 220 Issue 5 (05 03 2021) ISSN: 1540-8140 [Electronic] United States
• PMID: 33856408 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: © 2021 May et al.

Chemical References

• Hedgehog Proteins
• Intracellular Signaling Peptides and Proteins
• Receptors, G-Protein-Coupled
• protein kinase modulator
• Cyclic AMP
• Cyclic AMP-Dependent Protein Kinases

Topics

• Animals
• Cell Line
• Cilia (metabolism)
• Cyclic AMP (metabolism)
• Cyclic AMP-Dependent Protein Kinases (metabolism)
• Fibroblasts (metabolism)
• HEK293 Cells
• Hedgehog Proteins (metabolism)
• Humans
• Intracellular Signaling Peptides and Proteins (metabolism)
• Mice
• NIH 3T3 Cells
• Proteomics (methods)
• Receptors, G-Protein-Coupled (metabolism)
• Signal Transduction (physiology)