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Gαs is dispensable for β-arrestin coupling but dictates GRK selectivity and is predominant for gene expression regulation by β2-adrenergic receptor
β-arrestins play a key role in G protein–coupled receptor (GPCR) internalization, trafficking, and signaling. Whether β-arrestins act independently of G protein–mediated signaling has not been fully elucidated. Studies using genome-editing approaches revealed that whereas G proteins are essential fo...
Autores principales: | , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Biochemistry and Molecular Biology
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10641165/ https://www.ncbi.nlm.nih.gov/pubmed/37774973 http://dx.doi.org/10.1016/j.jbc.2023.105293 |
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author | Burghi, Valeria Paradis, Justine S. Officer, Adam Adame-Garcia, Sendi Rafael Wu, Xingyu Matthees, Edda S.F. Barsi-Rhyne, Benjamin Ramms, Dana J. Clubb, Lauren Acosta, Monica Tamayo, Pablo Bouvier, Michel Inoue, Asuka von Zastrow, Mark Hoffmann, Carsten Gutkind, J. Silvio |
author_facet | Burghi, Valeria Paradis, Justine S. Officer, Adam Adame-Garcia, Sendi Rafael Wu, Xingyu Matthees, Edda S.F. Barsi-Rhyne, Benjamin Ramms, Dana J. Clubb, Lauren Acosta, Monica Tamayo, Pablo Bouvier, Michel Inoue, Asuka von Zastrow, Mark Hoffmann, Carsten Gutkind, J. Silvio |
author_sort | Burghi, Valeria |
collection | PubMed |
description | β-arrestins play a key role in G protein–coupled receptor (GPCR) internalization, trafficking, and signaling. Whether β-arrestins act independently of G protein–mediated signaling has not been fully elucidated. Studies using genome-editing approaches revealed that whereas G proteins are essential for mitogen-activated protein kinase activation by GPCRs., β-arrestins play a more prominent role in signal compartmentalization. However, in the absence of G proteins, GPCRs may not activate β-arrestins, thereby limiting the ability to distinguish G protein from β-arrestin-mediated signaling events. We used β2-adrenergic receptor (β2AR) and its β2AR-C tail mutant expressed in human embryonic kidney 293 cells wildtype or CRISPR–Cas9 gene edited for Gα(s), β-arrestin1/2, or GPCR kinases 2/3/5/6 in combination with arrestin conformational sensors to elucidate the interplay between Gα(s) and β-arrestins in controlling gene expression. We found that Gα(s) is not required for β2AR and β-arrestin conformational changes, β-arrestin recruitment, and receptor internalization, but that Gα(s) dictates the GPCR kinase isoforms involved in β-arrestin recruitment. By RNA-Seq analysis, we found that protein kinase A and mitogen-activated protein kinase gene signatures were activated by stimulation of β2AR in wildtype and β-arrestin1/2-KO cells but absent in Gα(s)-KO cells. These results were validated by re-expressing Gα(s) in the corresponding KO cells and silencing β-arrestins in wildtype cells. These findings were extended to cellular systems expressing endogenous levels of β2AR. Overall, our results support that Gs is essential for β2AR-promoted protein kinase A and mitogen-activated protein kinase gene expression signatures, whereas β-arrestins initiate signaling events modulating Gα(s)-driven nuclear transcriptional activity. |
format | Online Article Text |
id | pubmed-10641165 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-106411652023-11-14 Gαs is dispensable for β-arrestin coupling but dictates GRK selectivity and is predominant for gene expression regulation by β2-adrenergic receptor Burghi, Valeria Paradis, Justine S. Officer, Adam Adame-Garcia, Sendi Rafael Wu, Xingyu Matthees, Edda S.F. Barsi-Rhyne, Benjamin Ramms, Dana J. Clubb, Lauren Acosta, Monica Tamayo, Pablo Bouvier, Michel Inoue, Asuka von Zastrow, Mark Hoffmann, Carsten Gutkind, J. Silvio J Biol Chem Research Article β-arrestins play a key role in G protein–coupled receptor (GPCR) internalization, trafficking, and signaling. Whether β-arrestins act independently of G protein–mediated signaling has not been fully elucidated. Studies using genome-editing approaches revealed that whereas G proteins are essential for mitogen-activated protein kinase activation by GPCRs., β-arrestins play a more prominent role in signal compartmentalization. However, in the absence of G proteins, GPCRs may not activate β-arrestins, thereby limiting the ability to distinguish G protein from β-arrestin-mediated signaling events. We used β2-adrenergic receptor (β2AR) and its β2AR-C tail mutant expressed in human embryonic kidney 293 cells wildtype or CRISPR–Cas9 gene edited for Gα(s), β-arrestin1/2, or GPCR kinases 2/3/5/6 in combination with arrestin conformational sensors to elucidate the interplay between Gα(s) and β-arrestins in controlling gene expression. We found that Gα(s) is not required for β2AR and β-arrestin conformational changes, β-arrestin recruitment, and receptor internalization, but that Gα(s) dictates the GPCR kinase isoforms involved in β-arrestin recruitment. By RNA-Seq analysis, we found that protein kinase A and mitogen-activated protein kinase gene signatures were activated by stimulation of β2AR in wildtype and β-arrestin1/2-KO cells but absent in Gα(s)-KO cells. These results were validated by re-expressing Gα(s) in the corresponding KO cells and silencing β-arrestins in wildtype cells. These findings were extended to cellular systems expressing endogenous levels of β2AR. Overall, our results support that Gs is essential for β2AR-promoted protein kinase A and mitogen-activated protein kinase gene expression signatures, whereas β-arrestins initiate signaling events modulating Gα(s)-driven nuclear transcriptional activity. American Society for Biochemistry and Molecular Biology 2023-09-27 /pmc/articles/PMC10641165/ /pubmed/37774973 http://dx.doi.org/10.1016/j.jbc.2023.105293 Text en © 2023 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Research Article Burghi, Valeria Paradis, Justine S. Officer, Adam Adame-Garcia, Sendi Rafael Wu, Xingyu Matthees, Edda S.F. Barsi-Rhyne, Benjamin Ramms, Dana J. Clubb, Lauren Acosta, Monica Tamayo, Pablo Bouvier, Michel Inoue, Asuka von Zastrow, Mark Hoffmann, Carsten Gutkind, J. Silvio Gαs is dispensable for β-arrestin coupling but dictates GRK selectivity and is predominant for gene expression regulation by β2-adrenergic receptor |
title | Gαs is dispensable for β-arrestin coupling but dictates GRK selectivity and is predominant for gene expression regulation by β2-adrenergic receptor |
title_full | Gαs is dispensable for β-arrestin coupling but dictates GRK selectivity and is predominant for gene expression regulation by β2-adrenergic receptor |
title_fullStr | Gαs is dispensable for β-arrestin coupling but dictates GRK selectivity and is predominant for gene expression regulation by β2-adrenergic receptor |
title_full_unstemmed | Gαs is dispensable for β-arrestin coupling but dictates GRK selectivity and is predominant for gene expression regulation by β2-adrenergic receptor |
title_short | Gαs is dispensable for β-arrestin coupling but dictates GRK selectivity and is predominant for gene expression regulation by β2-adrenergic receptor |
title_sort | gαs is dispensable for β-arrestin coupling but dictates grk selectivity and is predominant for gene expression regulation by β2-adrenergic receptor |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10641165/ https://www.ncbi.nlm.nih.gov/pubmed/37774973 http://dx.doi.org/10.1016/j.jbc.2023.105293 |
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