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PMID: 12937272 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Inhibition of proteasomal activity causes inclusion formation in neuronal and non-neuronal cells overexpressing Parkin.

Molecular biology of the cell ·Vol. 14 ·No. 11 ·2003-11-00 ·Pages 4541-56

Ardley HC, Scott GB, Rose SA, Tan NG, Markham AF, Robinson PA

Abstract

Association between protein inclusions and neurodegenerative diseases, including Parkinson's and Alzheimer's diseases, and polyglutamine disorders, has been widely documented. Although ubiquitin is conjugated to many of these aggregated proteins, the 26S proteasome does not efficiently degrade them. Mutations in the ubiquitin-protein ligase Parkin are associated with autosomal recessive juvenile Parkinsonism. Although Parkin-positive inclusions are not detected in brains of autosomal recessive juvenile Parkinsonism patients, Parkin is found in Lewy bodies in sporadic disease. This suggests that loss of Parkin ligase activity via mutation, or sequestration to Lewy bodies, is a contributory factor to sporadic disease onset. We now demonstrate that decreased proteasomal activity causes formation of large, noncytotoxic inclusions within the cytoplasm of both neuronal and nonneuronal cells overexpressing Parkin. This is not a general phenomenon as there is an absence of similar inclusions when HHARI, a structural homolog of Parkin, is overexpressed. The inclusions colocalize with ubiquitin and with proteasomes. Furthermore, Parkin inclusions colocalize with gamma-tubulin, acetylated alpha-tubulin, and cause redistribution of vimentin, suggesting aggresome-like properties. Our data imply that lower proteasomal activity, previously observed in brain tissue of Parkinson's disease patients, leads to Parkin accumulation and a concomitant reduction in ligase activity, thereby promoting Lewy body formation.

MeSH Terms
Animals COS Cells Carrier Proteins/genetics,metabolism Chlorocebus aethiops Cloning, Molecular Cysteine Endopeptidases/drug effects,genetics,metabolism Cysteine Proteinase Inhibitors/pharmacology Humans Hydrogen Peroxide/pharmacology Inclusion Bodies/enzymology,genetics Leupeptins/pharmacology Lewy Body Disease/genetics,metabolism Microscopy, Confocal Multienzyme Complexes/drug effects,genetics,metabolism Neurons/metabolism Osmotic Pressure/drug effects Oxidative Stress/drug effects Parkinson Disease/genetics,metabolism Proteasome Endopeptidase Complex Sorbitol/pharmacology Tubulin/metabolism Tumor Cells, Cultured Tunicamycin/pharmacology Ubiquitin/metabolism Ubiquitin-Protein Ligases/genetics,metabolism Vimentin/metabolism
Chemicals
Carrier Proteins Cysteine Proteinase Inhibitors Leupeptins Multienzyme Complexes Tubulin Ubiquitin Vimentin Tunicamycin Sorbitol Hydrogen Peroxide ARIH1 protein, human Ubiquitin-Protein Ligases parkin protein Cysteine Endopeptidases Proteasome Endopeptidase Complex benzyloxycarbonylleucyl-leucyl-leucine aldehyde
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Ardley Helen C
Molecular Medicine Unit, University of Leeds, St. James's University Hospital, Leeds LS9 7TF, United Kingdom. h.c.ardley@leeds.ac.uk
Scott Gina B
Rose Stephen A
Tan Nancy G S
Markham Alexander F
Robinson Philip A
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.e03-02-0078
Email
h.c.ardley@leeds.ac.uk
Published
2003-11-00
Epub
2003-00-22
Pages
4541-56
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC266771
Subset
IM
External Links
PubMed source DOI: 10.1091/mbc.e03-02-0078 PMC Full Text View journal details
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