title: FUsed in sarcoma is a novel regulator of manganese superoxide dismutase gene transcription.
abstract: OBJECTIVE
  FUsed in sarcoma (FUS) is a multifunctional DNA/RNA-binding protein that possesses diverse roles, such as RNA splicing, RNA transport, DNA repair, translation, and transcription. The network of enzymes and processes regulated by FUS is far from being fully described. In this study, we have focused on the mechanisms of FUS-regulated manganese superoxide dismutase (MnSOD) gene transcription.
  
  
  RESULTS
  Here we demonstrate that FUS is a component of the transcription complex that regulates the expression of MnSOD. Overexpression of FUS increased MnSOD expression in a dose-dependent manner and knockdown of FUS by siRNA led to the inhibition of MnSOD gene transcription. Reporter analyses, chromatin immunoprecipitation assay, electrophoretic mobility shift assay, affinity chromatography, and surface plasmon resonance analyses revealed the far upstream region of MnSOD promoter as an important target of FUS-mediated MnSOD transcription and confirmed that FUS binds to the MnSOD promoter and interacts with specificity protein 1 (Sp1). Importantly, overexpression of familial amyotropic lateral sclerosis (fALS)-linked R521G mutant FUS resulted in a significantly reduced level of MnSOD expression and activity, which is consistent with the decline in MnSOD activity observed in fibroblasts from fALS patients with the R521G mutation. R521G-mutant FUS abrogates MnSOD promoter-binding activity and interaction with Sp1.
  
  
  CONCLUSIONS
  This study identifies FUS as playing a critical role in MnSOD gene transcription and reveals a previously unrecognized relationship between MnSOD and mutant FUS in fALS.
authors.affiliation: 1 Graduate Center for Toxicology, University of Kentucky , Lexington, Kentucky.
authors.forename: Sanjit Kumar
authors.initials: SK
authors.lastname: Dhar
authors.forename: Jiayu
authors.initials: J
authors.lastname: Zhang
authors.forename: Jozsef
authors.initials: J
authors.lastname: Gal
authors.forename: Yong
authors.initials: Y
authors.lastname: Xu
authors.forename: Lu
authors.initials: L
authors.lastname: Miao
authors.forename: Bert C
authors.initials: BC
authors.lastname: Lynn
authors.forename: Haining
authors.initials: H
authors.lastname: Zhu
authors.forename: Edward J
authors.initials: EJ
authors.lastname: Kasarskis
authors.forename: Daret K
authors.initials: DK
authors.lastname: St Clair
mesh_terms: D000690:Amyotrophic Lateral Sclerosis / Q000201:enzymology / Q000235:genetics; D000818:Animals; D004790:Enzyme Induction; D056945:Hep G2 Cells; D006801:Humans; D007700:Kinetics; D051379:Mice; D009687:Nuclear Proteins / Q000378:metabolism; D000090243:Nucleophosmin; D017354:Point Mutation; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D034702:RNA-Binding Protein FUS / Q000737:chemistry / Q000502:physiology*; D016329:Sp1 Transcription Factor / Q000378:metabolism; D013482:Superoxide Dismutase / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D009687:Nuclear Proteins; D034702:RNA-Binding Protein FUS; D016329:Sp1 Transcription Factor; D000090243:Nucleophosmin; D013482:Superoxide Dismutase