title: The unfolded protein response modulates toxicity of the expanded glutamine androgen receptor.
abstract: Kennedy disease, a degenerative disorder caused by an expanded glutamine tract, is mediated by misfolding of the mutant androgen receptor (AR) protein, a process that may disrupt proteasome function. We hypothesized that this might lead to endoplasmic reticulum (ER) stress and induction of the unfolded protein response (UPR), a complex physiologic pathway that regulates cell survival. To test this hypothesis, we used aminoterminal fragments of wild type (AR16Q) or mutant (AR112Q) AR that triggered glutamine length-dependent cell death and activated an ER stress-inducible promoter. To evaluate the role of the UPR, we examined the contributions of three proximal sensors of ER stress: activating transcription factor 6 (ATF6), inositol requiring 1 (IRE1), and PKR-like endoplasmic reticulum kinase (PERK). AR112Q toxicity was significantly increased by a dominant negative ATF6 mutant and significantly decreased by a constitutively active ATF6 mutant, indicating that ATF6 promoted cell survival. In contrast, co-transfection with three separate IRE1alpha dominant negative mutants failed to alter glutamine length-dependent toxicity, suggesting that this arm of the UPR did not significantly affect AR112Q induced cell death. Activation of PERK, an ER transmembrane protein that functions as the third proximal UPR sensor, promoted glutamine length-dependent toxicity. Although nuclear localization sequence- and nuclear export sequence-targeted proteins both activated the UPR, this pathway more potently influenced toxicity when proteins were targeted to the cytoplasm. Taken together, our data demonstrate that the UPR is activated in cells expressing long glutamine tracts and that this pathway modulates polyglutamine toxicity.
authors.affiliation: Department of Pathology, the University of Michigan Medical School, Ann Arbor, Michigan 48109, USA.
authors.forename: Monzy
authors.initials: M
authors.lastname: Thomas
authors.forename: Zhigang
authors.initials: Z
authors.lastname: Yu
authors.forename: Nahid
authors.initials: N
authors.lastname: Dadgar
authors.forename: Sooryanarayana
authors.initials: S
authors.lastname: Varambally
authors.forename: Jianjun
authors.initials: J
authors.lastname: Yu
authors.forename: Arul M
authors.initials: AM
authors.lastname: Chinnaiyan
authors.forename: Andrew P
authors.initials: AP
authors.lastname: Lieberman
mesh_terms: D051702:Activating Transcription Factor 6; D021581:Active Transport, Cell Nucleus; D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D000818:Animals; D015153:Blotting, Western; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D016923:Cell Death; D002467:Cell Nucleus / Q000378:metabolism; D002470:Cell Survival; D003593:Cytoplasm / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005799:Genes, Dominant; D005973:Glutamine / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence; D009134:Muscular Atrophy, Spinal / Q000378:metabolism; D009154:Mutation; D041681:NIH 3T3 Cells; D020411:Oligonucleotide Array Sequence Analysis; D010455:Peptides / Q000737:chemistry / Q000633:toxicity*; D011401:Promoter Regions, Genetic; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D011489:Protein Denaturation; D017510:Protein Folding; D012313:RNA / Q000378:metabolism; D011944:Receptors, Androgen / Q000737:chemistry*; D013997:Time Factors; D014157:Transcription Factors / Q000378:metabolism; D014162:Transfection; D019892:eIF-2 Kinase / Q000378:metabolism
chemical_list: D051702:Activating Transcription Factor 6; D000590:Amino Acid Chloromethyl Ketones; D053148:Caspase 3; D020169:Caspases; D004268:DNA-Binding Proteins; D005973:Glutamine; D008156:Luciferases; D010455:Peptides; D046988:Proteasome Endopeptidase Complex; D012313:RNA; D011944:Receptors, Androgen; D014157:Transcription Factors; D019892:eIF-2 Kinase; C497111:ATF6 protein, human; C497112:Atf6 protein, mouse; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; C097188:polyglutamine; C505407:CASP3 protein, human; C505409:Casp3 protein, mouse