Ordered Phosphorylation Events in Two Independent Cascades of the PTEN C-tail Revealed by NMR
| Autor: | Elouan Terrien, Muriel Delepierre, Alain Chaffotte, Nicolas Wolff, Christophe Prehaud, François-Xavier Theillet, Henri Buc, Monique Lafon, Florence Cordier |
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| Přispěvatelé: | Résonance Magnétique Nucléaire des Biomolécules, Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], Cellule Pasteur UPMC, Institut Pasteur [Paris]-Sorbonne Université (SU), Neuro-Immunologie Virale, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris], This work was supported by the CNRS and grants from the Institut Pasteur, Agence Nationale pour la Recherche (PATHO-PDZ), and Institut Carnot Pasteur Maladies Infectieuses (NeuroVita). E.T. is a recipient of fellowships from the Ministère de l′Enseignement Supérieur et de la Recherche and from the Fondation pour la Recherche Médicale., We thank N. Kellershohn for valuable discussions, J. D’Alayer for help with mass spectrometry experiments and S. Baeriswyl for careful proofreading of the manuscript., ANR-06-MIME-0030,PATHO-PDZ,Role des proteines virales porteuses de site de fixation de domaine PDZ dans la pathogenicité virale. Analyse structure/fonction des complexes PDZ/PDZ-BS dans l'apoptose neuronale viro-induite.(2006), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP)-Sorbonne Université (SU), Institut Pasteur [Paris] (IP) |
| Jazyk: | angličtina |
| Rok vydání: | 2012 |
| Předmět: |
MESH: Glycogen Synthase Kinase 3 beta
Resonance structures [SDV]Life Sciences [q-bio] Peptides and proteins Biochemistry Catalysis law.invention 03 medical and health sciences Glycogen Synthase Kinase 3 0302 clinical medicine Colloid and Surface Chemistry GSK-3 law MESH: Nuclear Magnetic Resonance Biomolecular MESH: PTEN Phosphohydrolase / chemistry PTEN Humans [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology Phosphorylation MESH: Glycogen Synthase Kinase 3 / metabolism GSK3B Nuclear Magnetic Resonance Biomolecular MESH: PTEN Phosphohydrolase / metabolism 030304 developmental biology 0303 health sciences Glycogen Synthase Kinase 3 beta MESH: Humans biology Cluster chemistry MESH: Phosphorylation Chemistry Cell growth Kinase PTEN Phosphohydrolase General Chemistry 16. Peace & justice Cell biology [SDV.BBM.BP]Life Sciences [q-bio]/Biochemistry Molecular Biology/Biophysics 030220 oncology & carcinogenesis biology.protein Suppressor Cell and molecular biology Post-translational modification Intracellular |
| Zdroj: | Journal of the American Chemical Society Journal of the American Chemical Society, American Chemical Society, 2012, 134 (50), pp.20533-20543. ⟨10.1021/ja310214g⟩ Journal of the American Chemical Society, 2012, 134 (50), pp.20533-20543. ⟨10.1021/ja310214g⟩ |
| ISSN: | 0002-7863 1520-5126 |
| DOI: | 10.1021/ja310214g⟩ |
| Popis: | International audience; PTEN phosphatase is a tumor suppressor controlling notably cell growth, proliferation and survival. The multisite phosphorylation of the PTEN C-terminal tail regulates PTEN activity and intracellular trafficking. The dynamical nature of such regulatory events represents a crucial dimension for timing cellular decisions. Here we show that NMR spectroscopy allows reporting on the order and kinetics of clustered multisite phosphorylation events. We first unambiguously identify in vitro seven bona fide sites modified by CK2 and GSK3β kinases and two new sites on the PTEN C-terminal tail. Then, monitoring the formation of transient intermediate phosphorylated states, we determine the sequence of these reactions and calculate their apparent rate constants. Finally, we assess the dynamic formation of these phosphorylation events induced by endogenous kinases directly in extracts of human neuroblastoma cells. Taken together, our data indicate that two cascades of events controlled by CK2 and GSK3β occur independently on two clusters of sites (S380-S385 and S361-S370) and that in each cluster the reactions follow an ordered model with a distributive kinetic mechanism. Besides emphasizing the ability of NMR to quantitatively and dynamically follow post-translational modifications, these results bring a temporal dimension on the establishment of PTEN phosphorylation cascades. |
| Databáze: | OpenAIRE |
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