title: Sumoylation of the astroglial glutamate transporter EAAT2 governs its intracellular compartmentalization.
abstract: EAAT2 is a predominantly astroglial glutamate transporter responsible for the majority of synaptic glutamate clearance in the mammalian central nervous system (CNS). Its dysfunction has been linked with many neurological disorders, including amyotrophic lateral sclerosis (ALS). Decreases in EAAT2 expression and function have been implicated in causing motor neuron excitotoxic death in ALS. Nevertheless, increasing EAAT2 expression does not significantly improve ALS phenotype in mouse models or in clinical trials. In the SOD1-G93A mouse model of inherited ALS, the cytosolic carboxy-terminal domain is cleaved from EAAT2, conjugated to SUMO1, and accumulated in astrocytes where it triggers astrocyte-mediated neurotoxic effects as disease progresses. However, it is not known whether this fragment is sumoylated after cleavage or if full-length EAAT2 is already sumoylated prior to cleavage as part of physiological regulation. In this study, we show that a fraction of full-length EAAT2 is constitutively sumoylated in primary cultures of astrocytes in vitro and in the CNS in vivo. Furthermore, the extent of sumoylation of EAAT2 does not change during the course of ALS in the SOD1-G93A mouse and is not affected by the expression of ALS-causative mutant SOD1 proteins in astrocytes in vitro, indicating that EAAT2 sumoylation is not driven by pathogenic mechanisms. Most interestingly, sumoylated EAAT2 localizes to intracellular compartments, whereas non-sumoylated EAAT2 resides on the plasma membrane. In agreement, promoting desumoylation in primary astrocytes causes increased EAAT2-mediated glutamate uptake. These findings could have implications for optimizing therapeutic approaches aimed at increasing EAAT2 activity in the dysfunctional or diseased CNS.
authors.affiliation: Frances and Joseph Weinberg Unit for ALS Research, Farber Institute for Neuroscience, Department of Neuroscience, Thomas Jefferson University, Philadelphia, Pennsylvania.
authors.forename: E
authors.initials: E
authors.lastname: Foran
authors.forename: L
authors.initials: L
authors.lastname: Rosenblum
authors.forename: A
authors.initials: A
authors.lastname: Bogush
authors.forename: P
authors.initials: P
authors.lastname: Pasinelli
authors.forename: D
authors.initials: D
authors.lastname: Trotti
mesh_terms: D000690:Amyotrophic Lateral Sclerosis; D000818:Animals; D001253:Astrocytes / Q000378:metabolism*; D053148:Caspase 3 / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D004195:Disease Models, Animal; D027342:Excitatory Amino Acid Transporter 2 / Q000235:genetics / Q000378:metabolism*; D050491:Glutamate Plasma Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism*; D018698:Glutamic Acid / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism*; D008822:Mice, Transgenic; D016548:Prosencephalon / Q000378:metabolism; D017207:Rats, Sprague-Dawley; D013116:Spinal Cord / Q000378:metabolism; D058207:Sumoylation / Q000502:physiology*
chemical_list: D027342:Excitatory Amino Acid Transporter 2; D050491:Glutamate Plasma Membrane Transport Proteins; C493397:SLC1A2 protein, human; C493396:Slc1a2 protein, mouse; C493395:Slc1a2 protein, rat; D018698:Glutamic Acid; C505410:Casp3 protein, rat; D053148:Caspase 3
keywords: GLT-1; amyotrophic lateral sclerosis; excitotoxicity; protein trafficking
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