title: Molecular control of the amount, subcellular location, and activity state of translation elongation factor 2 in neurons experiencing stress.
abstract: Eukaryotic elongation factor 2 (eEF-2) is an important regulator of the protein translation machinery whereby it controls the movement of the ribosome along the mRNA. The activity of eEF-2 is regulated by changes in cellular energy status and nutrient availability and by posttranslational modifications such as phosphorylation and mono-ADP-ribosylation. However, the mechanisms regulating protein translation under conditions of cellular stress in neurons are unknown. Here we show that when rat hippocampal neurons experience oxidative stress (lipid peroxidation induced by exposure to cumene hydroperoxide; CH), eEF-2 is hyperphosphorylated and ribosylated, resulting in reduced translational activity. The degradation of eEF-2 requires calpain proteolytic activity and is accompanied by accumulation of eEF-2 in the nuclear compartment. The subcellular localization of both native and phosphorylated forms of eEF-2 is influenced by CRM1 and 14.3.3, respectively. In hippocampal neurons p53 interacts with nonphosphorylated (active) eEF-2, but not with its phosphorylated form. The p53-eEF-2 complexes are present in cytoplasm and nucleus, and their abundance increases when neurons experience oxidative stress. The nuclear localization of active eEF-2 depends upon its interaction with p53, as cells lacking p53 contain less active eEF-2 in the nuclear compartment. Overexpression of eEF-2 in hippocampal neurons results in increased nuclear levels of eEF-2 and decreased cell death after exposure to CH. Our results reveal novel molecular mechanisms controlling the differential subcellular localization and activity state of eEF-2 that may influence the survival status of neurons during periods of elevated oxidative stress.
authors.affiliation: Laboratory of Neurosciences, Intramural Research Program, National Institute on Aging, Baltimore, MD 21224, USA; Department of Biochemistry and Molecular Biology, Faculty of Pharmacy, University of Seville, Seville 41012, Spain.
authors.forename: Sandro
authors.initials: S
authors.lastname: Argüelles
authors.affiliation: Laboratory of Neurosciences, Intramural Research Program, National Institute on Aging, Baltimore, MD 21224, USA.
authors.forename: Simonetta
authors.initials: S
authors.lastname: Camandola
authors.affiliation: Laboratory of Neurosciences, Intramural Research Program, National Institute on Aging, Baltimore, MD 21224, USA; Department of Neuroscience, Brown University, Providence, RI 02912, USA.
authors.forename: Emmette R
authors.initials: ER
authors.lastname: Hutchison
authors.affiliation: Laboratory of Neurosciences, Intramural Research Program, National Institute on Aging, Baltimore, MD 21224, USA.
authors.forename: Roy G
authors.initials: RG
authors.lastname: Cutler
authors.affiliation: Department of Biochemistry and Molecular Biology, Faculty of Pharmacy, University of Seville, Seville 41012, Spain.
authors.forename: Antonio
authors.initials: A
authors.lastname: Ayala
authors.affiliation: Laboratory of Neurosciences, Intramural Research Program, National Institute on Aging, Baltimore, MD 21224, USA; Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. Electronic address: mattsonm@grc.nia.nih.gov.
authors.forename: Mark P
authors.initials: MP
authors.lastname: Mattson
mesh_terms: D048948:14-3-3 Proteins / Q000502:physiology; D000246:Adenosine Diphosphate Ribose / Q000378:metabolism; D000818:Animals; D001555:Benzene Derivatives / Q000494:pharmacology; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D045325:HCT116 Cells; D006801:Humans; D028884:Karyopherins / Q000502:physiology; D015227:Lipid Peroxidation; D009474:Neurons / Q000378:metabolism*; D018384:Oxidative Stress; D020652:Peptide Elongation Factor 2 / Q000032:analysis / Q000502:physiology*; D010766:Phosphorylation; D051381:Rats; D018160:Receptors, Cytoplasmic and Nuclear / Q000502:physiology; D016159:Tumor Suppressor Protein p53 / Q000502:physiology; D000097606:Exportin 1 Protein
chemical_list: D048948:14-3-3 Proteins; D000246:Adenosine Diphosphate Ribose; D001555:Benzene Derivatives; D028884:Karyopherins; D020652:Peptide Elongation Factor 2; D018160:Receptors, Cytoplasmic and Nuclear; D016159:Tumor Suppressor Protein p53; D000097606:Exportin 1 Protein; C007164:cumene hydroperoxide
keywords: 14.3.3; CRM1; Eukaryotic elongation factor 2; Free radicals; Hippocampal neurons; Lipid peroxidation; eEF-2; p53
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