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p62/SQSTM1-Dependent Autophagy of Lewy Body-Like α-Synuclein Inclusions
α-Synuclein is the main component of Lewy bodies, the intraneuronal inclusion bodies characteristic of Parkinson’s disease. Although α-synuclein accumulation is caused by inhibition of proteasome and autophagy-lysosome, the degradation of α-synuclein inclusions is still unknown. Formation of Lewy bo...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3534125/ https://www.ncbi.nlm.nih.gov/pubmed/23300799 http://dx.doi.org/10.1371/journal.pone.0052868 |
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author | Watanabe, Yoshihisa Tatebe, Harutsugu Taguchi, Katsutoshi Endo, Yasuhisa Tokuda, Takahiko Mizuno, Toshiki Nakagawa, Masanori Tanaka, Masaki |
author_facet | Watanabe, Yoshihisa Tatebe, Harutsugu Taguchi, Katsutoshi Endo, Yasuhisa Tokuda, Takahiko Mizuno, Toshiki Nakagawa, Masanori Tanaka, Masaki |
author_sort | Watanabe, Yoshihisa |
collection | PubMed |
description | α-Synuclein is the main component of Lewy bodies, the intraneuronal inclusion bodies characteristic of Parkinson’s disease. Although α-synuclein accumulation is caused by inhibition of proteasome and autophagy-lysosome, the degradation of α-synuclein inclusions is still unknown. Formation of Lewy body-like inclusions can be replicated in cultured cells by introducing α-synuclein fibrils generated in vitro. We used this cell culture model to investigate the autophagy of α-synuclein inclusions and impaired mitochondria. The intracellular α-synuclein inclusions immediately underwent phosphorylation and ubiquitination. Simultaneously they were encircled by an adaptor protein p62/SQSTM1 and directed to the autophagy-lysosome pathway in HEK293 cell line. Most phospho-α-synuclein-positive inclusions were degraded in 24 h, however, lysosomal dysfunction with bafilomycin A1 significantly affected their clearance. Moreover, inhibition of autophagy by Atg-5 siRNA treatment reduced the incorporation of α-synuclein inclusions into LC3-positive autophagosomes. Knockdown experiments demonstrated the requirement of p62 for α-synuclein autophagy. These results demonstrate that α-synuclein inclusions are preferred targets for p62-dependent autophagy. Next, we investigated the autophagic clearance of impaired mitochondria in α-synuclein inclusion-containing cells. Impaired mitochondria were almost completely eliminated after mitochondrial uncoupling even in the presence of α-synuclein inclusions, suggesting that mitochondrial clearance is not prevented by α-synuclein inclusions in HEK293 cells. |
format | Online Article Text |
id | pubmed-3534125 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-35341252013-01-08 p62/SQSTM1-Dependent Autophagy of Lewy Body-Like α-Synuclein Inclusions Watanabe, Yoshihisa Tatebe, Harutsugu Taguchi, Katsutoshi Endo, Yasuhisa Tokuda, Takahiko Mizuno, Toshiki Nakagawa, Masanori Tanaka, Masaki PLoS One Research Article α-Synuclein is the main component of Lewy bodies, the intraneuronal inclusion bodies characteristic of Parkinson’s disease. Although α-synuclein accumulation is caused by inhibition of proteasome and autophagy-lysosome, the degradation of α-synuclein inclusions is still unknown. Formation of Lewy body-like inclusions can be replicated in cultured cells by introducing α-synuclein fibrils generated in vitro. We used this cell culture model to investigate the autophagy of α-synuclein inclusions and impaired mitochondria. The intracellular α-synuclein inclusions immediately underwent phosphorylation and ubiquitination. Simultaneously they were encircled by an adaptor protein p62/SQSTM1 and directed to the autophagy-lysosome pathway in HEK293 cell line. Most phospho-α-synuclein-positive inclusions were degraded in 24 h, however, lysosomal dysfunction with bafilomycin A1 significantly affected their clearance. Moreover, inhibition of autophagy by Atg-5 siRNA treatment reduced the incorporation of α-synuclein inclusions into LC3-positive autophagosomes. Knockdown experiments demonstrated the requirement of p62 for α-synuclein autophagy. These results demonstrate that α-synuclein inclusions are preferred targets for p62-dependent autophagy. Next, we investigated the autophagic clearance of impaired mitochondria in α-synuclein inclusion-containing cells. Impaired mitochondria were almost completely eliminated after mitochondrial uncoupling even in the presence of α-synuclein inclusions, suggesting that mitochondrial clearance is not prevented by α-synuclein inclusions in HEK293 cells. Public Library of Science 2012-12-31 /pmc/articles/PMC3534125/ /pubmed/23300799 http://dx.doi.org/10.1371/journal.pone.0052868 Text en © 2012 Watanabe et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Watanabe, Yoshihisa Tatebe, Harutsugu Taguchi, Katsutoshi Endo, Yasuhisa Tokuda, Takahiko Mizuno, Toshiki Nakagawa, Masanori Tanaka, Masaki p62/SQSTM1-Dependent Autophagy of Lewy Body-Like α-Synuclein Inclusions |
title | p62/SQSTM1-Dependent Autophagy of Lewy Body-Like α-Synuclein Inclusions |
title_full | p62/SQSTM1-Dependent Autophagy of Lewy Body-Like α-Synuclein Inclusions |
title_fullStr | p62/SQSTM1-Dependent Autophagy of Lewy Body-Like α-Synuclein Inclusions |
title_full_unstemmed | p62/SQSTM1-Dependent Autophagy of Lewy Body-Like α-Synuclein Inclusions |
title_short | p62/SQSTM1-Dependent Autophagy of Lewy Body-Like α-Synuclein Inclusions |
title_sort | p62/sqstm1-dependent autophagy of lewy body-like α-synuclein inclusions |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3534125/ https://www.ncbi.nlm.nih.gov/pubmed/23300799 http://dx.doi.org/10.1371/journal.pone.0052868 |
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