title: A decrease in eukaryotic elongation factor 2 phosphorylation is required for local translation of sensorin and long-term facilitation in Aplysia.
abstract: Mechanistic target of rapamycin complex 1 (mTORC1)-dependent protein synthesis is required for many forms of synaptic plasticity and memory, but the downstream pathways important for synaptic plasticity are poorly understood. Long-term facilitation (LTF) in Aplysia is a form of synaptic plasticity that is closely linked to behavioral memory and an attractive model system for examining the important downstream targets for mTORC1 in regulating synaptic plasticity. Although mTORC1-regulated protein synthesis has been strongly linked to translation initiation, translation elongation is also regulated by mTORC1 and LTF leads to an mTORC1-dependent decrease in eukaryotic elongation factor 2 (eEF2) phosphorylation. The purpose of this study is to test the hypothesis that the decrease in eEF2 phosphorylation is required for mTORC1-dependent translation and plasticity. We show that the LTF-induced decrease in eEF2 phosphorylation is blocked by expression of an eEF2 kinase (eEF2K) modified to be resistant to mTORC1 regulation. We found that expression of this modified kinase blocked LTF. LTF requires local protein synthesis of the neuropeptide sensorin and importantly, local sensorin synthesis can be measured using a dendra fluorescent protein containing the 5' and 3' untranslated regions (UTRs) of sensorin. Using this construct, we show that blocking eEF2 dephosphorylation also blocks the increase in local sensorin synthesis. These results identify decreases in eEF2 phosphorylation as a critical downstream effector of mTOR required for long-term plasticity and identify an important translational target regulated by decreases in eEF2 phosphorylation.
authors.affiliation: Department of Neurology and Neurosurgery, Montreal Neurological Institute, McGill University, Montreal, Quebec, Canada.
authors.forename: Patrick K
authors.initials: PK
authors.lastname: McCamphill
authors.affiliation: Department of Neurology and Neurosurgery, Montreal Neurological Institute, McGill University, Montreal, Quebec, Canada.
authors.forename: Larissa
authors.initials: L
authors.lastname: Ferguson
authors.affiliation: Department of Neurology and Neurosurgery, Montreal Neurological Institute, McGill University, Montreal, Quebec, Canada.
authors.forename: Wayne S
authors.identifier: 0000-0003-1927-9315
authors.initials: WS
authors.lastname: Sossin
mesh_terms: D000818:Animals; D001048:Aplysia; D002478:Cells, Cultured; D054736:Elongation Factor 2 Kinase / Q000235:genetics / Q000378:metabolism*; D056890:Eukaryota / Q000378:metabolism*; D017774:Long-Term Potentiation / Q000502:physiology*; D009479:Neuropeptides / Q000378:metabolism; D020652:Peptide Elongation Factor 2 / Q000378:metabolism*; D010766:Phosphorylation; D015398:Signal Transduction / Q000502:physiology
chemical_list: D009479:Neuropeptides; D020652:Peptide Elongation Factor 2; D054736:Elongation Factor 2 Kinase
keywords: S6 kinase; eukaryotic elongation factor 2 kinase; learning and memory; protein synthesis; synaptic plasticity; target of rapamycin (TOR)