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Chaperone suppression of aggregation and altered subcellular proteasome localization imply protein misfolding in SCA1

Abstract

Spinocerebellar ataxia type 1 (SCA1) is an autosomal dominant neurodegenerative disorder caused by expansion of a polyglutamine tract in ataxin-1. In affected neurons of SCA1 patients and transgenic mice, mutant ataxin-1 accumulates in a single, ubiquitin-positive nuclear inclusion. In this study, we show that these inclusions stain positively for the 20S proteasome and the molecular chaperone HDJ-2/HSDJ. Similarly, HeLa cells transfected with mutant ataxin-1 develop nuclear aggregates which colocalize with the 20S proteasome and endogenous HDJ-2/HSDJ. Overexpression of wild-type HDJ-2/HSDJ in HeLa cells decreases the frequency of ataxin-1 aggregation. These data suggest that protein misfolding is responsible for the nuclear aggregates seen in SCA1, and that overexpression of a DnaJ chaperone promotes the recognition of a misfolded polyglutamine repeat protein, allowing its refolding and/or ubiquitin-dependent degradation.

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Figure 1: Immunohistochemical localization of 20S proteasome in brainstem neurons from an SCA1 patient and Purkinje cells of transgenic mice.
Figure 2: Immunohistochemical staining of HDJ-2/HSDJ in SCA1-patient neurons and transgenic mice Purkinje cells.
Figure 3: Ubiquitin immunostaining in COS7 cells expressing ataxin-1-GFP demonstrates the presence of ubiquitin in ataxin-1 aggregates.
Figure 4: Subcellular localization of 20S proteasome and ataxin-1 in HeLa cells.
Figure 5: Colocalization of endogenous HDJ-2/HSDJ and HSP70 with ataxin-1 nuclear aggregates.
Figure 6: Suppression of ataxin-1 aggregation in cells overexpressing HDJ-2/HSDJ.

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Acknowledgements

We thank W.F. Ward for the anti-proteasome antisera, G.N. DiMartino for anti-PA700 and anti-P31 antisera, J. K. Dunn for advice on statistical analyses, A.L. Beaudet, O.Lichtarge and W.E. O'Brien for their critical reading of the manuscript, and V. Brandt for her editorial help. This work is supported by grant from the National Institutes of Heath (NS27699 and NS22920) and by the Baylor Mental Retardation Research Center. H.Y.Z. is a Howard Hughes Medical Institute Investigator.

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Correspondence to Huda Y. Zoghbi.

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Cummings, C., Mancini, M., Antalffy, B. et al. Chaperone suppression of aggregation and altered subcellular proteasome localization imply protein misfolding in SCA1. Nat Genet 19, 148–154 (1998). https://doi.org/10.1038/502

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