Abstract
A dinucleotide deletion in human ubiquitin (Ub) B messenger RNA leads to formation of polyubiquitin (UbB)+1, which has been implicated in neuronal cell death in Alzheimer's and other neurodegenerative diseases. Previous studies demonstrate that UbB+1 protein causes proteasome dysfunction. However, the molecular mechanism of UbB+1-mediated neuronal degeneration remains unknown. We now report that UbB+1 causes neuritic beading, impairment of mitochondrial movements, mitochondrial stress and neuronal degeneration in primary neurons. Transfection of UbB+1 induced a buildup of mitochondria in neurites and dysregulation of mitochondrial motor proteins, in particular, through detachment of P74, the dynein intermediate chain, from mitochondria and decreased mitochondria–microtubule interactions. Altered distribution of mitochondria was associated with activation of both the mitochondrial stress and p53 cell death pathways. These results support the hypothesis that neuritic clogging of mitochondria by UbB+1 triggers a cascade of events characterized by local activation of mitochondrial stress followed by global cell death. Furthermore, UbB+1 small interfering RNA efficiently blocked expression of UbB+1 protein, attenuated neuritic beading and preserved cellular morphology, suggesting a potential neuroprotective strategy for certain neurodegenerative disorders.
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Abbreviations
- Ub:
-
ubiquitin
- UbB+1:
-
ubiquitin B+1
- AD:
-
Alzheimer's disease
- UPP:
-
uibiquitin-proteasome pathway
- siRNA:
-
small interfering RNA
- Aβ:
-
beta-amyloid
- CNS:
-
central nervous system
- MT:
-
microtubule
- EGFP:
-
enhanced green fluorescent protein
- EYFP:
-
enhanced yellow fluorescent protein
- ER:
-
endoplasmic reticulum
- TMRM:
-
tetramethyl rhodamine methyl ester
- MSR:
-
MitoSox Red
- TUNEL:
-
terminal dUTP nick-end labeling
- shRNA:
-
small hairpin RNA
- RFP:
-
red fluorescent protein
- GAPDH:
-
glyceraldehyde-3-phosphate dehydrogenase
- EM:
-
electron microscopy
- PCR:
-
DNA polymerase chain reaction
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Acknowledgements
This study was sponsored by NIH AG-26637 to Z.T., an Alzheimer's Association IIRG grant to S.S.S., NIH NS 27501 to D.K.O.D., NS 33213 to M.A.S., and grants from the Jan Dekker Stichting and Dr Ludgardine Bouwman Stichting (04–22), Internationale Stichting Alzheimer’s Onderzoek (04507/04830), the Van Leersum Foundation, the Hersenstichting Nederland (13F05.11) and the Hersenstichting Nederland-2005, the Matty Brand Stichting, the van Leersum Foundation (2006) and the Society for Progressive Supranuclear Palsy (440-4) to F.W.V.L.
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Tan, Z., Sun, X., Hou, FS. et al. Mutant ubiquitin found in Alzheimer's disease causes neuritic beading of mitochondria in association with neuronal degeneration. Cell Death Differ 14, 1721–1732 (2007). https://doi.org/10.1038/sj.cdd.4402180
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DOI: https://doi.org/10.1038/sj.cdd.4402180
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