title: IRE1 inhibition perturbs the unfolded protein response in a pancreatic β-cell line expressing mutant proinsulin, but does not sensitize the cells to apoptosis.
abstract: BACKGROUND
  The Akita mutation (C96Y) in the insulin gene results in early onset diabetes in both humans and mice. Expression of mutant proinsulin (C96Y) causes endoplasmic reticulum (ER) stress in pancreatic β-cells and consequently the cell activates the unfolded protein response (UPR). Since the proinsulin is terminally misfolded ER stress is irremediable and chronic activation of the UPR eventually activates apoptosis in some cells. Here we analyzed the IRE1-dependent activation of genes in response to misfolded proinsulin production in an inducible mutant proinsulin (C96Y) insulinoma cell line.
  
  
  RESULTS
  The IRE1 endoribonuclease inhibitors 4μ8c and MKC-3946 prevented the splicing of the XBP1 mRNA in response to ER stress caused by mutant proinsulin production. Microarray expression analysis and qPCR validation of select genes revealed that maximal upregulation of many UPR genes in response to mutant proinsulin production required IRE1, although most were still increased above control. Interestingly, neither degradation of misfolded proinsulin via ER-associated degradation (ERAD), nor apoptosis induced by prolonged misfolded proinsulin expression were affected by inhibiting IRE1.
  
  
  CONCLUSIONS
  Although maximal induction of most UPR genes requires IRE1, inhibition of IRE1 does not affect ERAD of misfolded proinsulin or predispose pancreatic β-cells expressing misfolded proinsulin to chronic ER stress-induced apoptosis.
authors.forename: Liling
authors.initials: L
authors.lastname: Zhang
authors.forename: Courtney
authors.initials: C
authors.lastname: Nosak
authors.forename: Pietro
authors.initials: P
authors.lastname: Sollazzo
authors.forename: Tanya
authors.initials: T
authors.lastname: Odisho
authors.affiliation: Division of Advanced Diagnostics-Metabolism, Toronto General Research Institute, University Health Network, 101 College Street, TMDT 10-706, Toronto, ON, Canada. allen.volchuk@utoronto.ca.
authors.forename: Allen
authors.initials: A
authors.lastname: Volchuk
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D045744:Cell Line, Tumor; D005786:Gene Expression Regulation / Q000187:drug effects; D050417:Insulin-Secreting Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D008565:Membrane Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D017354:Point Mutation; D011384:Proinsulin / Q000235:genetics* / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D059748:Proteolysis / Q000187:drug effects; D051381:Rats; D056811:Unfolded Protein Response / Q000187:drug effects*
chemical_list: D008565:Membrane Proteins; D047428:Protein Kinase Inhibitors; D011384:Proinsulin; C486557:Ern2 protein, rat; D017346:Protein Serine-Threonine Kinases