title: Genome wide array analysis indicates that an amyotrophic lateral sclerosis mutation of FUS causes an early increase of CAMK2N2 in vitro.
abstract: Mutations in the RNA binding protein FUS (fused in sarcoma) have been linked to a subset of familial amyotrophic lateral sclerosis (ALS) cases. The mutations are clustered in the C-terminal nuclear localization sequence (NLS). Various FUS mutants accumulate in the cytoplasm whereas wild-type (WT) FUS is mainly nuclear. Here we investigate the effect of one ALS causing mutant (FUS-ΔNLS, also known as R495X) on pre-mRNA splicing and RNA expression using genome wide exon-junction arrays. Using a non-neuronal stable cell line with inducible FUS expression, we detected early changes in RNA composition. In particular, mutant FUS-ΔNLS increased calcium/calmodulin-dependent protein kinase II inhibitor 2 (CAMK2N2) at both mRNA and protein levels, whereas WT-FUS had no effect. Chromatin immunoprecipitation experiments showed that FUS-ΔNLS accumulated at the CAMK2N2 promoter region, whereas promoter occupation by WT-FUS remained constant. Given the loss of FUS-ΔNLS in the nucleus through the mutation-induced translocation, this increase of promoter occupancy is surprising. It indicates that, despite the obvious cytoplasmic accumulation, FUS-ΔNLS can act through a nuclear gain of function mechanism.
authors.affiliation: Department of Molecular and Cellular Biochemistry, College of Medicine, University of Kentucky, 741 South Limestone, Lexington, KY 40536-0509, USA.
authors.forename: Paolo
authors.initials: P
authors.lastname: Convertini
authors.forename: Jiayu
authors.initials: J
authors.lastname: Zhang
authors.forename: Pierre
authors.initials: P
authors.lastname: de la Grange
authors.forename: Lawrence J
authors.initials: LJ
authors.lastname: Hayward
authors.forename: Haining
authors.initials: H
authors.lastname: Zhu
authors.forename: Stefan
authors.initials: S
authors.lastname: Stamm
mesh_terms: D000690:Amyotrophic Lateral Sclerosis / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2 / Q000096:biosynthesis / Q000235:genetics*; D002460:Cell Line; D002467:Cell Nucleus / Q000235:genetics; D002843:Chromatin / Q000235:genetics / Q000378:metabolism; D003593:Cytoplasm / Q000235:genetics; D005091:Exons; D055106:Genome-Wide Association Study / Q000379:methods; D057809:HEK293 Cells; D006801:Humans; D009154:Mutation; D011401:Promoter Regions, Genetic; D012322:RNA Precursors / Q000235:genetics; D012326:RNA Splicing / Q000235:genetics; D034702:RNA-Binding Protein FUS / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D002843:Chromatin; D012322:RNA Precursors; D034702:RNA-Binding Protein FUS; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2
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