title: A caspase-3-cleaved fragment of the glial glutamate transporter EAAT2 is sumoylated and targeted to promyelocytic leukemia nuclear bodies in mutant SOD1-linked amyotrophic lateral sclerosis.
abstract: EAAT2 (excitatory amino acid transporter 2) is a high affinity, Na+-dependent glutamate transporter of glial origin that is essential for the clearance of synaptically released glutamate and prevention of excitotoxicity. During the course of human amyotrophic lateral sclerosis (ALS) and in a transgenic mutant SOD1 mouse model of the disease, expression and activity of EAAT2 is remarkably reduced. We previously showed that some of the mutant SOD1 proteins exposed to oxidative stress inhibit EAAT2 by triggering caspase-3 cleavage of EAAT2 at a single defined locus. This gives rise to two fragments that we termed truncated EAAT2 and COOH terminus of EAAT2 (CTE). In this study, we report that analysis of spinal cord homogenates prepared from mutant G93A-SOD1 mice reveals CTE to be of a higher molecular weight than expected because it is conjugated with SUMO-1. The sumoylated CTE fragment (CTE-SUMO-1) accumulates in the spinal cord of these mice as early as presymptomatic stage (70 days of age) and not in other central nervous system areas unaffected by the disease. The presence and accumulation of CTE-SUMO-1 is specific to ALS mice, since it does not occur in the R6/2 mouse model for Huntington disease. Furthermore, using an astroglial cell line, primary culture of astrocytes, and tissue samples from G93A-SOD1 mice, we show that CTE-SUMO-1 is targeted to promyelocytic leukemia nuclear bodies. Since one of the proposed functions of promyelocytic leukemia nuclear bodies is regulation of gene transcription, we suggest a possible novel mechanism by which the glial glutamate transporter EAAT2 could contribute to the pathology of ALS.
authors.affiliation: Farber Institute for Neurosciences, Weinberg Unit for ALS Research, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, and Cecil B. Day Laboratory for Neuromuscular Research, Massachusetts General Hospital, Charlestown 02129, USA.
authors.forename: Stuart L
authors.initials: SL
authors.lastname: Gibb
authors.forename: William
authors.initials: W
authors.lastname: Boston-Howes
authors.forename: Zeno S
authors.initials: ZS
authors.lastname: Lavina
authors.forename: Stefano
authors.initials: S
authors.lastname: Gustincich
authors.forename: Robert H
authors.initials: RH
authors.lastname: Brown
authors.forename: Piera
authors.initials: P
authors.lastname: Pasinelli
authors.forename: Davide
authors.initials: D
authors.lastname: Trotti
mesh_terms: D000690:Amyotrophic Lateral Sclerosis / Q000473:pathology*; D000818:Animals; D030801:Animals, Genetically Modified; D001253:Astrocytes / Q000378:metabolism; D053148:Caspase 3 / Q000378:metabolism*; D022003:Cell Nucleus Structures / Q000378:metabolism; D027342:Excitatory Amino Acid Transporter 2 / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D006816:Huntington Disease / Q000473:pathology; D047468:Immunoprecipitation; D045586:Intranuclear Inclusion Bodies / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D008970:Molecular Weight; D017434:Protein Structure, Tertiary; D051381:Rats; D025842:SUMO-1 Protein / Q000378:metabolism*; D013116:Spinal Cord / Q000473:pathology; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism*; D000072105:Superoxide Dismutase-1