title: Dual oxidase maturation factor 1 (DUOXA1) overexpression increases reactive oxygen species production and inhibits murine muscle satellite cell differentiation.
abstract: BACKGROUND
  Dual oxidase maturation factor 1 (DUOXA1) has been associated with the maturation of the reactive oxygen species (ROS) producing enzyme, dual oxidase 1 (DUOX1) in the adult thyroid. However, ROS have also been implicated in the development of several tissues. We found that activated muscle satellite cells and primary myoblasts isolated from mice express robust levels of DUOXA1 and that its levels are altered as cells differentiate.
  
  
  RESULTS
  To determine whether DUOXA1 levels affect muscle differentiation, we used an adenoviral construct (pCMV5-DUOXA1-GFP) to drive constitutive overexpression of this protein in primary myoblasts. High levels of DUOXA1 throughout myogenesis resulted in enhanced H2O2 production, fusion defects, reduced expression of early (myogenin) and late (myosin heavy chain) markers of differentiation, and elevated levels of apoptosis compared to control cells infected with an empty adenoviral vector (pCMV5-GFP). DUOXA1 knockdown (using a DUOXA1 shRNA construct) resulted in enhanced differentiation compared to cells subjected to a control shRNA, and subjecting DUOXA1 overexpressing cells to siRNAs targeting DUOX1 or apoptosis signal-regulating kinase 1 (ASK1) rescued the phenotype.
  
  
  CONCLUSIONS
  This study represents the first to demonstrate the importance of DUOXA1 in skeletal muscle myoblasts and that DUOXA1 overexpression in muscle stem cells induces apoptosis and inhibits differentiation through DUOX1 and ASK1.
authors.affiliation: Siebens-Drake Research Institute, 1400 Western Road, London, Ontario N6G 2 V4, Canada. sdsandiford@gmail.com.
authors.forename: Shelley D E
authors.initials: SD
authors.lastname: Sandiford
authors.forename: Karen A M
authors.initials: KA
authors.lastname: Kennedy
authors.forename: Xiaojun
authors.initials: X
authors.lastname: Xie
authors.forename: J Geoffrey
authors.initials: JG
authors.lastname: Pickering
authors.forename: Shawn S C
authors.initials: SS
authors.lastname: Li
mesh_terms: D000818:Animals; D017209:Apoptosis; D002454:Cell Differentiation; D002478:Cells, Cultured; D000074623:Dual Oxidases; D048848:MAP Kinase Kinase Kinase 5 / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D032446:Myoblasts / Q000166:cytology / Q000378:metabolism*; D019255:NADPH Oxidases / Q000235:genetics / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D032496:Satellite Cells, Skeletal Muscle / Q000166:cytology / Q000378:metabolism*
chemical_list: C560186:DUOXA1 protein, mouse; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; D017382:Reactive Oxygen Species; D000074623:Dual Oxidases; D019255:NADPH Oxidases; C519130:Duox1 protein, mouse; D048848:MAP Kinase Kinase Kinase 5; C482859:Map3k5 protein, mouse