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Extended polyglutamine selectively interacts with caspase-8 and -10 in nuclear aggregates
1Department of Genetics, National Children's Medical Research Center, 3-35-31, Taishido, Setagaya, Tokyo 154-8509, Japan.
Abstract:
A growing number of inherited neurodegenerative disorders, including Huntington's disease, have been shown to be caused by the expansion of CAG/polyglutamine repeats. The molecular mechanism underlying these disorders, however, has yet to be clarified. We and others previously demonstrated that caspase-8 was activated by proteolysis in association with the expression of extended polyglutamine. Here, we further analyzed the selectivity of caspases in the process mediated by extended polyglutamine. Among upstream caspases, caspase-10, a close homolog of caspase-8, was also proteolytically activated, but caspase-9 was not. Caspase-8 and -10 were recruited into nuclear aggregates of extended polyglutamine, where at least a fraction of these caspases was converted to the activated forms. Caspase-8 and -10 were co-immunoprecipitated with polyglutamine only when the polyglutamine was pathologically extended, whereas caspase-2, -3, -6, -7 and -9 were not co-immunoprecipitated with polyglutamine regardless of its size. A dominant-negative form of caspase-8 with a mutation at the catalytic cysteine residue inhibited polyglutamine-mediated nuclear apoptotic phenotype. These results suggest that caspase-8 and -10 are autoactivated as a result of close proximity of the proforms of these molecules that occurs due to aggregate formation, which reveals a novel toxic gain-of-function mechanism for the pathogenesis of CAG-repeat disorders.
Insights
Extended polyglutamine in neurodegenerative diseases activates caspase-8 and -10. This aggregate-induced autoactivation reveals a novel toxic gain-of-function mechanism in CAG-repeat disorders.
Area of Science:
- Neuroscience
- Molecular Biology
- Genetics
Background:
- Inherited neurodegenerative disorders like Huntington's disease are linked to expanded CAG/polyglutamine repeats.
- The precise molecular mechanisms driving these conditions remain unclear.
Purpose of the Study:
- To investigate the role and selectivity of caspases in polyglutamine-mediated pathogenesis.
- To elucidate the mechanism of caspase activation in the context of expanded polyglutamine.
Main Methods:
- Analysis of caspase activation and recruitment to nuclear polyglutamine aggregates.
- Co-immunoprecipitation assays to determine caspase-polyglutamine interactions.
- Functional studies using dominant-negative caspase-8 to assess its role in polyglutamine-induced apoptosis.
Main Results:
- Caspase-8 and its homolog caspase-10 were proteolytically activated and recruited to nuclear polyglutamine aggregates.
- Caspase-8 and -10, but not other caspases, co-immunoprecipitated with pathologically extended polyglutamine.
- Inhibition of caspase-8 activity blocked polyglutamine-mediated nuclear apoptotic phenotypes.
Conclusions:
- Caspase-8 and -10 autoactivate due to proximity within polyglutamine aggregates.
- This represents a novel toxic gain-of-function mechanism contributing to CAG-repeat disorder pathogenesis.