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Developmental regulation of p70 S6 kinase by a G protein-coupled receptor dynamically modelized in primary cells

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Abstract

The mechanisms whereby G protein-coupled receptors (GPCR) activate signalling pathways involved in mRNA translation are ill-defined, in contrast to tyrosine kinase receptors (TKR). We compared a GPCR and a TKR, both endogenously expressed, for their ability to mediate phosphorylation of 70-kDa ribosomal S6 kinase p70S6K in primary rat Sertoli cells at two developmental stages. In proliferating cells stimulated with follicle-stimulating hormone (FSH), active p70S6K was phosphorylated on T389 and T421/S424, through cAMP-dependent kinase (PKA) and phosphatidyl-inositide-3 kinase (PI3K) antagonizing actions. In FSH-stimulated differentiating cells, active p70S6K was phosphorylated solely on T389, PKA and PI3K independently enhancing its activity. At both developmental stages, insulin-induced p70S6K regulation was consistent with reported data. Therefore, TKR and GPCR trigger distinct p70S6K active conformations. p70S6K developmental regulation was formalized in a dynamic mathematical model fitting the data, which led to experimentally inaccessible predictions on p70S6K phosphorylation rate.

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Acknowledgments

The authors thank the technical assistance of the rat breeders Jean-Claude Braguer and Claude Cahier. Drs Florian Guillou, Laurence Dupuy in our laboratory, and Drs François Fages and Frédérique Clément (INRIA, Rocquencourt), are acknowledged for critical reading of the manuscript. We are also indebted to Drs Len Stephens and Phil Hawkins for the PIP3 quantification and to Dr Anne Segonds-Pichon for statistical analysis (Babraham Institute, Cambridge, UK). A.M. was funded by a fellowship from the Region Centre and from the Institut National de la Recherche Agronomique (INRA), D.H. was funded by a fellowship from the INRA (ASC), G.D. by a fellowship from the Ministère de la Recherche et de la Technologie, C.L. was granted by the Fondation pour le Recherche Médicale, and H.G. was a BBSRC fellow. This work was supported by the INRA AgroBI AIP, by the Centre National de la Recherche Scientifique, by Université de Tours and by the AE INRIA/INRA REGATE.

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Authors and Affiliations

  1. BIOS Group, INRA, UMR85, Unité Physiologie de la Reproduction et des Comportements, 37380, Nouzilly, France

    Astrid Musnier, Domitille Heitzler, Thomas Boulo, Guillaume Durand, Anne Poupon, Eric Reiter & Pascale Crépieux

  2. CNRS, UMR6175, 37380, Nouzilly, France

    Astrid Musnier, Domitille Heitzler, Thomas Boulo, Guillaume Durand, Anne Poupon, Eric Reiter & Pascale Crépieux

  3. Université François Rabelais, 37041, Tours, France

    Astrid Musnier, Domitille Heitzler, Thomas Boulo, Guillaume Durand, Anne Poupon, Eric Reiter & Pascale Crépieux

  4. Haras Nationaux, 37380, Nouzilly, France

    Astrid Musnier, Domitille Heitzler, Thomas Boulo, Guillaume Durand, Anne Poupon, Eric Reiter & Pascale Crépieux

  5. INRA UR83 de Recherches Avicoles, 37380, Nouzilly, France

    Sophie Tesseraud

  6. The Inositide Laboratory, The Babraham Institute, Cambridge, CB2 4AT, UK

    Charlotte Lécureuil & Hervé Guillou

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  1. Astrid Musnier
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  2. Domitille Heitzler
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Correspondence to Pascale Crépieux.

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Musnier, A., Heitzler, D., Boulo, T. et al. Developmental regulation of p70 S6 kinase by a G protein-coupled receptor dynamically modelized in primary cells. Cell. Mol. Life Sci. 66, 3487–3503 (2009). https://doi.org/10.1007/s00018-009-0134-z

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