1. GPR161 structure uncovers the redundant role of sterol-regulated ciliary cAMP signaling in the Hedgehog pathway.
- Author
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Hoppe, Nicholas, Harrison, Simone, Hwang, Sun-Hee, Chen, Ziwei, Karelina, Masha, Deshpande, Ishan, Suomivuori, Carl-Mikael, Palicharla, Vivek, Berry, Samuel, Tschaikner, Philipp, Regele, Dominik, Covey, Douglas, Stefan, Eduard, Marks, Debora, Reiter, Jeremy, Dror, Ron, Evers, Alex, Mukhopadhyay, Saikat, and Manglik, Aashish
- Subjects
Humans ,Hedgehog Proteins ,Signal Transduction ,Receptors ,G-Protein-Coupled ,Mutation ,Cilia - Abstract
The orphan G protein-coupled receptor (GPCR) GPR161 plays a central role in development by suppressing Hedgehog signaling. The fundamental basis of how GPR161 is activated remains unclear. Here, we determined a cryogenic-electron microscopy structure of active human GPR161 bound to heterotrimeric Gs. This structure revealed an extracellular loop 2 that occupies the canonical GPCR orthosteric ligand pocket. Furthermore, a sterol that binds adjacent to transmembrane helices 6 and 7 stabilizes a GPR161 conformation required for Gs coupling. Mutations that prevent sterol binding to GPR161 suppress Gs-mediated signaling. These mutants retain the ability to suppress GLI2 transcription factor accumulation in primary cilia, a key function of ciliary GPR161. By contrast, a protein kinase A-binding site in the GPR161 C terminus is critical in suppressing GLI2 ciliary accumulation. Our work highlights how structural features of GPR161 interface with the Hedgehog pathway and sets a foundation to understand the role of GPR161 function in other signaling pathways.
- Published
- 2024