title: PKA-dependent phosphorylation of ribosomal protein S6 does not correlate with translation efficiency in striatonigral and striatopallidal medium-sized spiny neurons.
abstract: Ribosomal protein S6 (rpS6), a component of the 40S ribosomal subunit, is phosphorylated on several residues in response to numerous stimuli. Although commonly used as a marker for neuronal activity, its upstream mechanisms of regulation are poorly studied and its role in protein synthesis remains largely debated. Here, we demonstrate that the psychostimulant d-amphetamine (d-amph) markedly increases rpS6 phosphorylation at Ser235/236 sites in both crude and synaptoneurosomal preparations of the mouse striatum. This effect occurs selectively in D1R-expressing medium-sized spiny neurons (MSNs) and requires the cAMP/PKA/DARPP-32/PP-1 cascade, whereas it is independent of mTORC1/p70S6K, PKC, and ERK signaling. By developing a novel assay to label nascent peptidic chains, we show that the rpS6 phosphorylation induced in striatonigral MSNs by d-amph, as well as in striatopallidal MSNs by the antipsychotic haloperidol or in both subtypes by papaverine, is not correlated with the translation of global or 5' terminal oligopyrimidine tract mRNAs. Together, these results provide novel mechanistic insights into the in vivo regulation of the post-translational modification of rpS6 in the striatum and point out the lack of a relationship between PKA-dependent rpS6 phosphorylation and translation efficiency.
authors.affiliation: Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5203, Institut de Génomique Fonctionnelle, Montpellier, F-34094, France, INSERM, U1191, Montpellier, F-34094, France; Université de Montpellier, Montpellier, F-34094, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Biever
authors.affiliation: Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5203, Institut de Génomique Fonctionnelle, Montpellier, F-34094, France, INSERM, U1191, Montpellier, F-34094, France; Université de Montpellier, Montpellier, F-34094, France.
authors.forename: Emma
authors.initials: E
authors.lastname: Puighermanal
authors.affiliation: Department of Pharmacology, Kurume University School of Medicine, Kurume, Fukuoka 830-0011, Japan.
authors.forename: Akinori
authors.initials: A
authors.lastname: Nishi
authors.affiliation: Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5203, Institut de Génomique Fonctionnelle, Montpellier, F-34094, France, INSERM, U1191, Montpellier, F-34094, France; Université de Montpellier, Montpellier, F-34094, France.
authors.forename: Alexandre
authors.initials: A
authors.lastname: David
authors.affiliation: Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5203, Institut de Génomique Fonctionnelle, Montpellier, F-34094, France, INSERM, U1191, Montpellier, F-34094, France; Université de Montpellier, Montpellier, F-34094, France.
authors.forename: Claire
authors.initials: C
authors.lastname: Panciatici
authors.affiliation: Université Pierre et Marie Curie, INSERM UMR-S839, Paris, F-75005 France, Institut du Fer à Moulin, F-75005, Paris, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Longueville
authors.affiliation: Centre de Recherche en Biochimie Macromoléculaire, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5237, Montpellier, F-34000, France, and.
authors.forename: Dimitris
authors.initials: D
authors.lastname: Xirodimas
authors.affiliation: Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5203, Institut de Génomique Fonctionnelle, Montpellier, F-34094, France, INSERM, U1191, Montpellier, F-34094, France; Université de Montpellier, Montpellier, F-34094, France.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Gangarossa
authors.affiliation: Department of Biochemistry and Molecular Biology, The Institute for Medical Research Israel-Canada, The Hebrew University-Hadassah Medical School, Jerusalem 91120, Israel.
authors.forename: Oded
authors.initials: O
authors.lastname: Meyuhas
authors.affiliation: Université Pierre et Marie Curie, INSERM UMR-S839, Paris, F-75005 France, Institut du Fer à Moulin, F-75005, Paris, France.
authors.forename: Denis
authors.initials: D
authors.lastname: Hervé
authors.affiliation: Université Pierre et Marie Curie, INSERM UMR-S839, Paris, F-75005 France, Institut du Fer à Moulin, F-75005, Paris, France.
authors.forename: Jean-Antoine
authors.initials: JA
authors.lastname: Girault
authors.affiliation: Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5203, Institut de Génomique Fonctionnelle, Montpellier, F-34094, France, INSERM, U1191, Montpellier, F-34094, France; Université de Montpellier, Montpellier, F-34094, France, emmanuel.valjent@igf.cnrs.fr emmanuel.valjent@gmail.com.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Valjent
mesh_terms: D000818:Animals; D003342:Corpus Striatum / Q000166:cytology* / Q000187:drug effects; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D051937:Dopamine and cAMP-Regulated Phosphoprotein 32 / Q000235:genetics / Q000378:metabolism; D005260:Female; D044385:GTP-Binding Protein alpha Subunits / Q000235:genetics / Q000378:metabolism; D006248:Harringtonines / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009434:Neural Pathways / Q000187:drug effects / Q000502:physiology*; D009474:Neurons / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D010766:Phosphorylation / Q000187:drug effects / Q000235:genetics; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D021381:Protein Transport / Q000187:drug effects / Q000235:genetics; D011691:Puromycin / Q000494:pharmacology; D017447:Receptors, Dopamine D1 / Q000235:genetics / Q000378:metabolism; D017448:Receptors, Dopamine D2 / Q000235:genetics / Q000378:metabolism; D038601:Ribosomal Protein S6 / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D013378:Substantia Nigra / Q000166:cytology* / Q000187:drug effects; D013574:Synaptosomes / Q000187:drug effects / Q000378:metabolism
chemical_list: D051937:Dopamine and cAMP-Regulated Phosphoprotein 32; C583261:Drd1 protein, mouse; D044385:GTP-Binding Protein alpha Subunits; D006248:Harringtonines; C497929:Ppp1r1b protein, mouse; D011500:Protein Synthesis Inhibitors; D017447:Receptors, Dopamine D1; D017448:Receptors, Dopamine D2; D038601:Ribosomal Protein S6; C084618:olfactory G protein subunit alpha olf; D011691:Puromycin; D017868:Cyclic AMP-Dependent Protein Kinases
keywords: d-amphetamine; dopamine; protein synthesis; ribosomal protein S6; striatum