Regulation of the RNA-binding protein Smaug by the GPCR Smoothened via the kinase Fused

Lucia Bruzzone; Camilla Argüelles; Matthieu Sanial; Samia Miled; Giorgia Alvisi; Marina Gonçalves-Antunes; Fairouz Qasrawi; Robert A. Holmgren; Craig A. Smibert; Howard D. Lipshitz; Graciela L. Boccaccio; Anne Plessis; Isabelle Bécam

doi:10.15252/embr.201948425

Regulation of the RNA-binding protein Smaug by the GPCR Smoothened via the kinase Fused

Lucia Bruzzone, Camilla Argüelles, Matthieu Sanial^*, Samia Miled, Giorgia Alvisi, Marina Gonçalves-Antunes, Fairouz Qasrawi, Robert A. Holmgren, Craig A. Smibert, Howard D. Lipshitz, Graciela L. Boccaccio, Anne Plessis^*, Isabelle Bécam^*

^*Corresponding author for this work

Molecular Biosciences

Research output: Contribution to journal › Article › peer-review

8 Scopus citations

Abstract

From fly to mammals, the Smaug/Samd4 family of prion-like RNA-binding proteins control gene expression by destabilizing and/or repressing the translation of numerous target transcripts. However, the regulation of its activity remains poorly understood. We show that Smaug's protein levels and mRNA repressive activity are downregulated by Hedgehog signaling in tissue culture cells. These effects rely on the interaction of Smaug with the G-protein coupled receptor Smoothened, which promotes the phosphorylation of Smaug by recruiting the kinase Fused. The activation of Fused and its binding to Smaug are sufficient to suppress its ability to form cytosolic bodies and to antagonize its negative effects on endogenous targets. Importantly, we demonstrate in vivo that HH reduces the levels of smaug mRNA and increases the level of several mRNAs downregulated by Smaug. Finally, we show that Smaug acts as a positive regulator of Hedgehog signaling during wing morphogenesis. These data constitute the first evidence for a post-translational regulation of Smaug and reveal that the fate of several mRNAs bound to Smaug is modulated by a major signaling pathway.

Original language	English (US)
Article number	e48425
Journal	EMBO Reports
Volume	21
Issue number	7
DOIs	https://doi.org/10.15252/embr.201948425
State	Published - Jul 3 2020

Funding

We thank late Dr. E. Izaurralde for her help with the tethering assay Drs. D. Hipfner, J.Jia. J. Jiang, M. Simonelig, and P. Therond and their respective laboratories for plasmids and antibodies; Dr. F. Besse and L. Ruel for their invaluable advices. We are grateful to G. Baldacci, V. Courtier, V. Doye, A. Guichet, M. Nadal L. Pintard, F. Schweisguth, M. Wener, and our colleagues from the IJM and the fly community for their support and insightful discussions and to R. Cervouze for his technical support. Antibodies from the DSHB were developed under the auspices of the NICHD and maintained by the University of Iowa. embryo injections were carried out by BestGene Inc. We thank the ImagoSeine imaging Facility which was funded by the ARC, the region Ile de France (SESAME), and Paris‐Diderot University. Paris‐Diderot University (ARS) and CNRS jointly funded the LC‐MS/MS equipment of the Proteomics Facility of the Institute Jacques Monod and the Region Ile‐de‐France (SESAME). This work was supported by the CNRS, the University of Paris, the Fondation ARC pour la Recherche sur le Cancer (grant 1112), and ECOSSud. C. A. C. A. was supported by l'Association Franco‐argentine, by the Fondation ARC pour la Recherche sur le Cancer, and by an international fellowship from CONICET‐Argentina, and L.B. by Paris Sorbonne Cité. , Drosophila We thank late Dr. E. Izaurralde for her help with the tethering assay, Drs. D. Hipfner, J.Jia. J. Jiang, M. Simonelig, and P. Therond and their respective laboratories for plasmids and antibodies; Dr. F. Besse and L. Ruel for their invaluable advices. We are grateful to G. Baldacci, V. Courtier, V. Doye, A. Guichet, M. Nadal L. Pintard, F. Schweisguth, M. Wener, and our colleagues from the IJM and the fly community for their support and insightful discussions and to R. Cervouze for his technical support. Antibodies from the DSHB were developed under the auspices of the NICHD and maintained by the University of Iowa. Drosophila embryo injections were carried out by BestGene Inc. We thank the ImagoSeine imaging Facility which was funded by the ARC, the region Ile de France (SESAME), and Paris-Diderot University. Paris-Diderot University (ARS) and CNRS jointly funded the LC-MS/MS equipment of the Proteomics Facility of the Institute Jacques Monod and the Region Ile-de-France (SESAME). This work was supported by the CNRS, the University of Paris, the Fondation ARC pour la Recherche sur le Cancer (grant 1112), and ECOSSud. C. A. C. A. was supported by l'Association Franco-argentine, by the Fondation ARC pour la Recherche sur le Cancer, and by an international fellowship from CONICET-Argentina, and L.B. by Paris Sorbonne Cit?.

Keywords

Drosophila
Hedgehog
SAMD4
Smaug
Smoothened

ASJC Scopus subject areas

Genetics
Molecular Biology
Biochemistry

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10.15252/embr.201948425

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title = "Regulation of the RNA-binding protein Smaug by the GPCR Smoothened via the kinase Fused",

abstract = "From fly to mammals, the Smaug/Samd4 family of prion-like RNA-binding proteins control gene expression by destabilizing and/or repressing the translation of numerous target transcripts. However, the regulation of its activity remains poorly understood. We show that Smaug's protein levels and mRNA repressive activity are downregulated by Hedgehog signaling in tissue culture cells. These effects rely on the interaction of Smaug with the G-protein coupled receptor Smoothened, which promotes the phosphorylation of Smaug by recruiting the kinase Fused. The activation of Fused and its binding to Smaug are sufficient to suppress its ability to form cytosolic bodies and to antagonize its negative effects on endogenous targets. Importantly, we demonstrate in vivo that HH reduces the levels of smaug mRNA and increases the level of several mRNAs downregulated by Smaug. Finally, we show that Smaug acts as a positive regulator of Hedgehog signaling during wing morphogenesis. These data constitute the first evidence for a post-translational regulation of Smaug and reveal that the fate of several mRNAs bound to Smaug is modulated by a major signaling pathway.",

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author = "Lucia Bruzzone and Camilla Arg{\"u}elles and Matthieu Sanial and Samia Miled and Giorgia Alvisi and Marina Gon{\c c}alves-Antunes and Fairouz Qasrawi and Holmgren, {Robert A.} and Smibert, {Craig A.} and Lipshitz, {Howard D.} and Boccaccio, {Graciela L.} and Anne Plessis and Isabelle B{\'e}cam",

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T1 - Regulation of the RNA-binding protein Smaug by the GPCR Smoothened via the kinase Fused

AU - Bruzzone, Lucia

AU - Argüelles, Camilla

AU - Sanial, Matthieu

AU - Miled, Samia

AU - Alvisi, Giorgia

AU - Gonçalves-Antunes, Marina

AU - Qasrawi, Fairouz

AU - Holmgren, Robert A.

AU - Smibert, Craig A.

AU - Lipshitz, Howard D.

AU - Boccaccio, Graciela L.

AU - Plessis, Anne

AU - Bécam, Isabelle

PY - 2020/7/3

Y1 - 2020/7/3

N2 - From fly to mammals, the Smaug/Samd4 family of prion-like RNA-binding proteins control gene expression by destabilizing and/or repressing the translation of numerous target transcripts. However, the regulation of its activity remains poorly understood. We show that Smaug's protein levels and mRNA repressive activity are downregulated by Hedgehog signaling in tissue culture cells. These effects rely on the interaction of Smaug with the G-protein coupled receptor Smoothened, which promotes the phosphorylation of Smaug by recruiting the kinase Fused. The activation of Fused and its binding to Smaug are sufficient to suppress its ability to form cytosolic bodies and to antagonize its negative effects on endogenous targets. Importantly, we demonstrate in vivo that HH reduces the levels of smaug mRNA and increases the level of several mRNAs downregulated by Smaug. Finally, we show that Smaug acts as a positive regulator of Hedgehog signaling during wing morphogenesis. These data constitute the first evidence for a post-translational regulation of Smaug and reveal that the fate of several mRNAs bound to Smaug is modulated by a major signaling pathway.

AB - From fly to mammals, the Smaug/Samd4 family of prion-like RNA-binding proteins control gene expression by destabilizing and/or repressing the translation of numerous target transcripts. However, the regulation of its activity remains poorly understood. We show that Smaug's protein levels and mRNA repressive activity are downregulated by Hedgehog signaling in tissue culture cells. These effects rely on the interaction of Smaug with the G-protein coupled receptor Smoothened, which promotes the phosphorylation of Smaug by recruiting the kinase Fused. The activation of Fused and its binding to Smaug are sufficient to suppress its ability to form cytosolic bodies and to antagonize its negative effects on endogenous targets. Importantly, we demonstrate in vivo that HH reduces the levels of smaug mRNA and increases the level of several mRNAs downregulated by Smaug. Finally, we show that Smaug acts as a positive regulator of Hedgehog signaling during wing morphogenesis. These data constitute the first evidence for a post-translational regulation of Smaug and reveal that the fate of several mRNAs bound to Smaug is modulated by a major signaling pathway.

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KW - Smoothened

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Regulation of the RNA-binding protein Smaug by the GPCR Smoothened via the kinase Fused

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