title: Induction of HSP70 expression and recruitment of HSC70 and HSP70 in the nucleus reduce aggregation of a polyalanine expansion mutant of PABPN1 in HeLa cells.
abstract: Nuclear inclusions formed by the aggregation of a polyalanine expansion mutant of the nuclear poly(A)-binding protein (PABPN1) is a hallmark of oculopharyngeal muscular dystrophy (OPMD). OPMD is a dominant autosomal disease in which patients exhibit progressive difficulty of swallowing and eyelid elevation, starting around the age of 50. At present, there is no specific treatment to reduce the aggregate burden in patients. However, in cell culture models of OPMD, reduction of protein aggregation can be achieved by ectopic expression of HSP70. As gene transfer may not be the most effective means to elevate HSP70 levels, we tested four pharmacological agents for their ability to induce HSP70, recruit both HSP70 and HSC70 into the cell nucleus and reduce mutant PABPN1 aggregation in a HeLa cell culture model. We show here that exposure to moderate levels of ZnSO4, 8-hydroxyquinoline, ibuprofen and indomethacin produced a robust stress response resulting in the induction of HSP70 in HeLa cells expressing the mutant PABPN1 as a green fluorescent protein (GFP) fusion protein. Both HSP70 and the constitutive chaperone HSC70 localized in the nucleus of cells treated with any one of the four agents. This stress response was similar to what was observed following hyperthermia. All four agents also caused a significant reduction in the cellular burden of protein aggregates, as was judged by confocal microscopy and solubility changes of the aggregates. A concomitant reduction of cell death in drug-treated mutant PABPN1 expressing cells was also observed.
authors.affiliation: Department of Molecular and Cellular Biology, University of Guelph, Ont, Canada.
authors.forename: Qishan
authors.initials: Q
authors.lastname: Wang
authors.forename: Dick D
authors.initials: DD
authors.lastname: Mosser
authors.forename: Jnanankur
authors.initials: J
authors.lastname: Bag
mesh_terms: D021581:Active Transport, Cell Nucleus; D002467:Cell Nucleus / Q000737:chemistry / Q000378:metabolism; D049452:Green Fluorescent Proteins; D050883:HSC70 Heat-Shock Proteins / Q000032:analysis / Q000378:metabolism*; D018840:HSP70 Heat-Shock Proteins / Q000032:analysis / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D007052:Ibuprofen / Q000494:pharmacology; D007213:Indomethacin / Q000494:pharmacology; D045586:Intranuclear Inclusion Bodies / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D039141:Muscular Dystrophy, Oculopharyngeal / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D017830:Octoxynol / Q000737:chemistry; D015125:Oxyquinoline / Q000494:pharmacology; D010455:Peptides / Q000235:genetics*; D039122:Poly(A)-Binding Protein I / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D061988:Proteasome Inhibitors; D012995:Solubility; D014162:Transfection; D015854:Up-Regulation; D019287:Zinc Sulfate / Q000494:pharmacology
chemical_list: D050883:HSC70 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; D010455:Peptides; D039122:Poly(A)-Binding Protein I; D061988:Proteasome Inhibitors; D049452:Green Fluorescent Proteins; C019529:polyalanine; D015125:Oxyquinoline; D019287:Zinc Sulfate; D017830:Octoxynol; D046988:Proteasome Endopeptidase Complex; D007052:Ibuprofen; D007213:Indomethacin