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GPR161 structure uncovers the redundant role of sterol-regulated ciliary cAMP signaling in the Hedgehog pathway
The orphan G protein-coupled receptor (GPCR) GPR161 is enriched in primary cilia, where it plays a central role in suppressing Hedgehog signaling(1). GPR161 mutations lead to developmental defects and cancers(2,3,4). The fundamental basis of how GPR161 is activated, including potential endogenous ac...
Autores principales: | , , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cold Spring Harbor Laboratory
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10245861/ https://www.ncbi.nlm.nih.gov/pubmed/37292845 http://dx.doi.org/10.1101/2023.05.23.540554 |
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author | Hoppe, Nicholas Harrison, Simone Hwang, Sun-Hee Chen, Ziwei Karelina, Masha Deshpande, Ishan Suomivuori, Carl-Mikael Palicharla, Vivek R. Berry, Samuel P. Tschaikner, Philipp Regele, Dominik Covey, Douglas F. Stefan, Eduard Marks, Debora S. Reiter, Jeremy Dror, Ron O. Evers, Alex S. Mukhopadhyay, Saikat Manglik, Aashish |
author_facet | Hoppe, Nicholas Harrison, Simone Hwang, Sun-Hee Chen, Ziwei Karelina, Masha Deshpande, Ishan Suomivuori, Carl-Mikael Palicharla, Vivek R. Berry, Samuel P. Tschaikner, Philipp Regele, Dominik Covey, Douglas F. Stefan, Eduard Marks, Debora S. Reiter, Jeremy Dror, Ron O. Evers, Alex S. Mukhopadhyay, Saikat Manglik, Aashish |
author_sort | Hoppe, Nicholas |
collection | PubMed |
description | The orphan G protein-coupled receptor (GPCR) GPR161 is enriched in primary cilia, where it plays a central role in suppressing Hedgehog signaling(1). GPR161 mutations lead to developmental defects and cancers(2,3,4). The fundamental basis of how GPR161 is activated, including potential endogenous activators and pathway-relevant signal transducers, remains unclear. To elucidate GPR161 function, we determined a cryogenic-electron microscopy structure of active GPR161 bound to the heterotrimeric G protein complex G(s). This structure revealed an extracellular loop 2 that occupies the canonical GPCR orthosteric ligand pocket. Furthermore, we identify a sterol that binds to a conserved extrahelical site adjacent to transmembrane helices 6 and 7 and stabilizes a GPR161 conformation required for G(s) coupling. Mutations that prevent sterol binding to GPR161 suppress cAMP pathway activation. Surprisingly, these mutants retain the ability to suppress GLI2 transcription factor accumulation in cilia, a key function of ciliary GPR161 in Hedgehog pathway suppression. By contrast, a protein kinase A-binding site in the GPR161 C-terminus is critical in suppressing GLI2 ciliary accumulation. Our work highlights how unique structural features of GPR161 interface with the Hedgehog pathway and sets a foundation to understand the broader role of GPR161 function in other signaling pathways. |
format | Online Article Text |
id | pubmed-10245861 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Cold Spring Harbor Laboratory |
record_format | MEDLINE/PubMed |
spelling | pubmed-102458612023-06-08 GPR161 structure uncovers the redundant role of sterol-regulated ciliary cAMP signaling in the Hedgehog pathway Hoppe, Nicholas Harrison, Simone Hwang, Sun-Hee Chen, Ziwei Karelina, Masha Deshpande, Ishan Suomivuori, Carl-Mikael Palicharla, Vivek R. Berry, Samuel P. Tschaikner, Philipp Regele, Dominik Covey, Douglas F. Stefan, Eduard Marks, Debora S. Reiter, Jeremy Dror, Ron O. Evers, Alex S. Mukhopadhyay, Saikat Manglik, Aashish bioRxiv Article The orphan G protein-coupled receptor (GPCR) GPR161 is enriched in primary cilia, where it plays a central role in suppressing Hedgehog signaling(1). GPR161 mutations lead to developmental defects and cancers(2,3,4). The fundamental basis of how GPR161 is activated, including potential endogenous activators and pathway-relevant signal transducers, remains unclear. To elucidate GPR161 function, we determined a cryogenic-electron microscopy structure of active GPR161 bound to the heterotrimeric G protein complex G(s). This structure revealed an extracellular loop 2 that occupies the canonical GPCR orthosteric ligand pocket. Furthermore, we identify a sterol that binds to a conserved extrahelical site adjacent to transmembrane helices 6 and 7 and stabilizes a GPR161 conformation required for G(s) coupling. Mutations that prevent sterol binding to GPR161 suppress cAMP pathway activation. Surprisingly, these mutants retain the ability to suppress GLI2 transcription factor accumulation in cilia, a key function of ciliary GPR161 in Hedgehog pathway suppression. By contrast, a protein kinase A-binding site in the GPR161 C-terminus is critical in suppressing GLI2 ciliary accumulation. Our work highlights how unique structural features of GPR161 interface with the Hedgehog pathway and sets a foundation to understand the broader role of GPR161 function in other signaling pathways. Cold Spring Harbor Laboratory 2023-05-24 /pmc/articles/PMC10245861/ /pubmed/37292845 http://dx.doi.org/10.1101/2023.05.23.540554 Text en https://creativecommons.org/licenses/by-nc-nd/4.0/This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which allows reusers to copy and distribute the material in any medium or format in unadapted form only, for noncommercial purposes only, and only so long as attribution is given to the creator. |
spellingShingle | Article Hoppe, Nicholas Harrison, Simone Hwang, Sun-Hee Chen, Ziwei Karelina, Masha Deshpande, Ishan Suomivuori, Carl-Mikael Palicharla, Vivek R. Berry, Samuel P. Tschaikner, Philipp Regele, Dominik Covey, Douglas F. Stefan, Eduard Marks, Debora S. Reiter, Jeremy Dror, Ron O. Evers, Alex S. Mukhopadhyay, Saikat Manglik, Aashish GPR161 structure uncovers the redundant role of sterol-regulated ciliary cAMP signaling in the Hedgehog pathway |
title | GPR161 structure uncovers the redundant role of sterol-regulated ciliary cAMP signaling in the Hedgehog pathway |
title_full | GPR161 structure uncovers the redundant role of sterol-regulated ciliary cAMP signaling in the Hedgehog pathway |
title_fullStr | GPR161 structure uncovers the redundant role of sterol-regulated ciliary cAMP signaling in the Hedgehog pathway |
title_full_unstemmed | GPR161 structure uncovers the redundant role of sterol-regulated ciliary cAMP signaling in the Hedgehog pathway |
title_short | GPR161 structure uncovers the redundant role of sterol-regulated ciliary cAMP signaling in the Hedgehog pathway |
title_sort | gpr161 structure uncovers the redundant role of sterol-regulated ciliary camp signaling in the hedgehog pathway |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10245861/ https://www.ncbi.nlm.nih.gov/pubmed/37292845 http://dx.doi.org/10.1101/2023.05.23.540554 |
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