Several missense mutations in the protein kinase Cγ (γPKC) gene have been found to cause spinocerebellar ataxia type 14 (SCA14), an autosomal dominant neurodegenerative disease. We previously demonstrated that the mutant γPKC found in SCA14 is misfolded, susceptible to aggregation and cytotoxic. Molecular chaperones assist the refolding and degradation of misfolded proteins and prevention of the proteins' aggregation. In the present study, we found that the expression of mutant γPKC-GFP increased the levels of heat-shock protein 70 (Hsp70) in SH-SY5Y cells. To elucidate the role of this elevation, we investigated the effect of siRNA-mediated knockdown of Hsp70 on the aggregation and cytotoxicity of mutant γPKC. Knockdown of Hsp70 exacerbated the aggregation and cytotoxicity of mutant γPKC-GFP by inhibiting this mutant's degradation. These findings suggest that mutant γPKC increases the level of Hsp70, which protects cells from the mutant's cytotoxicity by enhancing its degradation.
Keywords: 17-AAG; 17-allylamino-17-demethoxygeldanamycin; Aggregation; CHIP; Cytotoxicity; GFP; Heat-shock protein 70; Hsc70; Hsp40; Hsp70; Hsp90; JNK; LDH; PCs; Purkinje cells; SCA14; Spinocerebellar ataxia; WT; c-Jun N-terminal kinase; carboxyl terminus of Hsp70-interacting protein; green fluorescent protein; heat-shock cognate protein 70; heat-shock protein 40; heat-shock protein 70; heat-shock protein 90; lactate dehydrogenase; protein kinase Cγ; spinocerebellar ataxia type 14; wild-type; γPKC.
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