title: Small heat-shock proteins interact with a flanking domain to suppress polyglutamine aggregation.
abstract: Small heat-shock proteins (sHsps) are molecular chaperones that play an important protective role against cellular protein misfolding by interacting with partially unfolded proteins on their off-folding pathway, preventing their aggregation. Polyglutamine (polyQ) repeat expansion leads to the formation of fibrillar protein aggregates and neuronal cell death in nine diseases, including Huntington disease and the spinocerebellar ataxias (SCAs). There is evidence that sHsps have a role in suppression of polyQ-induced neurodegeneration; for example, the sHsp alphaB-crystallin (alphaB-c) has been identified as a suppressor of SCA3 toxicity in a Drosophila model. However, the molecular mechanism for this suppression is unknown. In this study we tested the ability of alphaB-c to suppress the aggregation of a polyQ protein. We found that alphaB-c does not inhibit the formation of SDS-insoluble polyQ fibrils. We further tested the effect of alphaB-c on the aggregation of ataxin-3, a polyQ protein that aggregates via a two-stage aggregation mechanism. The first stage involves association of the N-terminal Josephin domain followed by polyQ-mediated interactions and the formation of SDS-resistant mature fibrils. Our data show that alphaB-c potently inhibits the first stage of ataxin-3 aggregation; however, the second polyQ-dependent stage can still proceed. By using NMR spectroscopy, we have determined that alphaB-c interacts with an extensive region on the surface of the Josephin domain. These data provide an example of a domain/region flanking an amyloidogenic sequence that has a critical role in modulating aggregation of a polypeptide and plays a role in the interaction with molecular chaperones to prevent this aggregation.
authors.affiliation: Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, 3800, Australia.
authors.forename: Amy L
authors.initials: AL
authors.lastname: Robertson
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Headey
authors.forename: Helen M
authors.initials: HM
authors.lastname: Saunders
authors.forename: Heath
authors.initials: H
authors.lastname: Ecroyd
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Scanlon
authors.forename: John A
authors.initials: JA
authors.lastname: Carver
authors.forename: Stephen P
authors.initials: SP
authors.lastname: Bottomley
mesh_terms: D050888:Heat-Shock Proteins, Small / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D046529:Microscopy, Electron, Transmission; D008958:Models, Molecular; D009419:Nerve Tissue Proteins / Q000737:chemistry; D019906:Nuclear Magnetic Resonance, Biomolecular; D010455:Peptides / Q000737:chemistry*; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D012995:Solubility; D038203:alpha-Crystallin B Chain / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure
chemical_list: D050888:Heat-Shock Proteins, Small; D009419:Nerve Tissue Proteins; D010455:Peptides; D038203:alpha-Crystallin B Chain; C097188:polyglutamine