title: Altered expression of HSPA5, HSPA8 and PARK7 in spinocerebellar ataxia type 17 identified by 2-dimensional fluorescence difference in gel electrophoresis.
abstract: BACKGROUND
  Expansion of the CAG repeat of the TATA-box binding protein (TBP) gene has been identified as the causative mutations in spinocerebellar ataxia 17 (SCA17). TBP is ubiquitously expressed in both central nervous system and peripheral tissues. The underlying molecular changes of SCA17 are rarely explored.
  
  
  METHODS
  To study the molecular mechanisms underlying SCA17, we generated stably induced isogenic 293 cells expressing normal TBP-Q(36) and expanded TBP-Q(61) and analyzed the expressed proteins using two-dimensional difference in gel electrophoresis (2D-DIGE), followed by mass spectrometry and immunoblotting.
  
  
  RESULTS
  Upon induction with doxycycline, the expanded TBP-Q(61) formed aggregates with significant increase in the cell population at subG1 phase and cleaved caspase-3. Proteomics study identified a total of 16 proteins with expression changes greater than 1.5 fold. Among the 16 proteins, PARK7, GLRX3, HNRNPA1, GINS1, ENO1, HNRPK and NPM1 are increased, and SERPINA5, HSPA5, VCL, KHSRP, HSPA8, HNRPH1, IMMT, VCP and HNRNPL are decreased in cells expressing TBP-Q(61) compared with those expressing TBP-Q(36). The altered expression of HSPA5, HSPA8 and PARK7 were further validated by 2D and Western immunoblot analyses.
  
  
  CONCLUSIONS
  The results illustrate the utility of proteomics to identify alterations of proteins which underlie pathogenesis of SCA17, and may serve as potential therapeutic targets.
authors.affiliation: Department of Life Science, National Taiwan Normal University, Taipei, Taiwan.
authors.forename: Li-Ching
authors.initials: LC
authors.lastname: Lee
authors.forename: Chiung-Mei
authors.initials: CM
authors.lastname: Chen
authors.forename: Fen-Lin
authors.initials: FL
authors.lastname: Chen
authors.forename: Pei-Ying
authors.initials: PY
authors.lastname: Lin
authors.forename: Ya-Chin
authors.initials: YC
authors.lastname: Hsiao
authors.forename: Pin-Rong
authors.initials: PR
authors.lastname: Wang
authors.forename: Ming-Tsan
authors.initials: MT
authors.lastname: Su
authors.forename: Hsiu-Mei
authors.initials: HM
authors.lastname: Hsieh-Li
authors.forename: Ji-Chuu
authors.initials: JC
authors.lastname: Hwang
authors.forename: Chung-Hsin
authors.initials: CH
authors.lastname: Wu
authors.forename: Guan-Chiun
authors.initials: GC
authors.lastname: Lee
authors.forename: Sher
authors.initials: S
authors.lastname: Singh
authors.forename: Yenshou
authors.initials: Y
authors.lastname: Lin
authors.forename: Sen-Yung
authors.initials: SY
authors.lastname: Hsieh
authors.forename: Guey-Jen
authors.initials: GJ
authors.lastname: Lee-Chen
authors.forename: Jung-Yaw
authors.initials: JY
authors.lastname: Lin
mesh_terms: D015153:Blotting, Western; D002460:Cell Line; D003001:Cloning, Molecular; D015180:Electrophoresis, Gel, Two-Dimensional; D000091342:Endoplasmic Reticulum Chaperone BiP; D005453:Fluorescence; D005786:Gene Expression Regulation; D050883:HSC70 Heat-Shock Proteins / Q000032:analysis* / Q000235:genetics; D006360:Heat-Shock Proteins / Q000032:analysis* / Q000235:genetics; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000032:analysis* / Q000235:genetics; D018832:Molecular Chaperones / Q000032:analysis* / Q000235:genetics; D000090243:Nucleophosmin; D015513:Oncogene Proteins / Q000032:analysis* / Q000235:genetics; D010641:Phenotype; D000071617:Protein Deglycase DJ-1; D020754:Spinocerebellar Ataxias / Q000235:genetics* / Q000378:metabolism*; D035181:TATA-Box Binding Protein / Q000235:genetics
chemical_list: D000091342:Endoplasmic Reticulum Chaperone BiP; D050883:HSC70 Heat-Shock Proteins; C000717247:HSPA5 protein, human; C494371:HSPA8 protein, human; D006360:Heat-Shock Proteins; D047908:Intracellular Signaling Peptides and Proteins; D018832:Molecular Chaperones; C000716967:NPM1 protein, human; D015513:Oncogene Proteins; D035181:TATA-Box Binding Protein; D000090243:Nucleophosmin; C105131:PARK7 protein, human; D000071617:Protein Deglycase DJ-1