Cargando…

Amyloid-Like Aggregates of the Yeast Prion Protein Ure2 Enter Vertebrate Cells by Specific Endocytotic Pathways and Induce Apoptosis

BACKGROUND: A number of amyloid diseases involve deposition of extracellular protein aggregates, which are implicated in mechanisms of cell damage and death. However, the mechanisms involved remain poorly understood. METHODOLOGY/PRINCIPAL FINDINGS: Here we use the yeast prion protein Ure2 as a gener...

Descripción completa

Detalles Bibliográficos
Autores principales: Zhang, Chen, Jackson, Antony P., Zhang, Zai-Rong, Han, Yan, Yu, Shun, He, Rong-Qiao, Perrett, Sarah
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2932714/
https://www.ncbi.nlm.nih.gov/pubmed/20824085
http://dx.doi.org/10.1371/journal.pone.0012529
_version_ 1782186093096992768
author Zhang, Chen
Jackson, Antony P.
Zhang, Zai-Rong
Han, Yan
Yu, Shun
He, Rong-Qiao
Perrett, Sarah
author_facet Zhang, Chen
Jackson, Antony P.
Zhang, Zai-Rong
Han, Yan
Yu, Shun
He, Rong-Qiao
Perrett, Sarah
author_sort Zhang, Chen
collection PubMed
description BACKGROUND: A number of amyloid diseases involve deposition of extracellular protein aggregates, which are implicated in mechanisms of cell damage and death. However, the mechanisms involved remain poorly understood. METHODOLOGY/PRINCIPAL FINDINGS: Here we use the yeast prion protein Ure2 as a generic model to investigate how amyloid-like protein aggregates can enter mammalian cells and convey cytotoxicity. The effect of three different states of Ure2 protein (native dimer, protofibrils and mature fibrils) was tested on four mammalian cell lines (SH-SY5Y, MES23.5, HEK-293 and HeLa) when added extracellularly to the medium. Immunofluorescence using a polyclonal antibody against Ure2 showed that all three protein states could enter the four cell lines. In each case, protofibrils significantly inhibited the growth of the cells in a dose-dependent manner, fibrils showed less toxicity than protofibrils, while the native state had no effect on cell growth. This suggests that the structural differences between the three protein states lead to their different effects upon cells. Protofibrils of Ure2 increased membrane conductivity, altered calcium homeostasis, and ultimately induced apoptosis. The use of standard inhibitors suggested uptake into mammalian cells might occur via receptor-mediated endocytosis. In order to investigate this further, we used the chicken DT40 B cell line DKOR, which allows conditional expression of clathrin. Uptake into the DKOR cell-line was reduced when clathrin expression was repressed suggesting similarities between the mechanism of PrP uptake and the mechanism observed here for Ure2. CONCLUSIONS/SIGNIFICANCE: The results provide insight into the mechanisms by which amyloid aggregates may cause pathological effects in prion and amyloid diseases.
format Text
id pubmed-2932714
institution National Center for Biotechnology Information
language English
publishDate 2010
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-29327142010-09-07 Amyloid-Like Aggregates of the Yeast Prion Protein Ure2 Enter Vertebrate Cells by Specific Endocytotic Pathways and Induce Apoptosis Zhang, Chen Jackson, Antony P. Zhang, Zai-Rong Han, Yan Yu, Shun He, Rong-Qiao Perrett, Sarah PLoS One Research Article BACKGROUND: A number of amyloid diseases involve deposition of extracellular protein aggregates, which are implicated in mechanisms of cell damage and death. However, the mechanisms involved remain poorly understood. METHODOLOGY/PRINCIPAL FINDINGS: Here we use the yeast prion protein Ure2 as a generic model to investigate how amyloid-like protein aggregates can enter mammalian cells and convey cytotoxicity. The effect of three different states of Ure2 protein (native dimer, protofibrils and mature fibrils) was tested on four mammalian cell lines (SH-SY5Y, MES23.5, HEK-293 and HeLa) when added extracellularly to the medium. Immunofluorescence using a polyclonal antibody against Ure2 showed that all three protein states could enter the four cell lines. In each case, protofibrils significantly inhibited the growth of the cells in a dose-dependent manner, fibrils showed less toxicity than protofibrils, while the native state had no effect on cell growth. This suggests that the structural differences between the three protein states lead to their different effects upon cells. Protofibrils of Ure2 increased membrane conductivity, altered calcium homeostasis, and ultimately induced apoptosis. The use of standard inhibitors suggested uptake into mammalian cells might occur via receptor-mediated endocytosis. In order to investigate this further, we used the chicken DT40 B cell line DKOR, which allows conditional expression of clathrin. Uptake into the DKOR cell-line was reduced when clathrin expression was repressed suggesting similarities between the mechanism of PrP uptake and the mechanism observed here for Ure2. CONCLUSIONS/SIGNIFICANCE: The results provide insight into the mechanisms by which amyloid aggregates may cause pathological effects in prion and amyloid diseases. Public Library of Science 2010-09-02 /pmc/articles/PMC2932714/ /pubmed/20824085 http://dx.doi.org/10.1371/journal.pone.0012529 Text en Zhang 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
Zhang, Chen
Jackson, Antony P.
Zhang, Zai-Rong
Han, Yan
Yu, Shun
He, Rong-Qiao
Perrett, Sarah
Amyloid-Like Aggregates of the Yeast Prion Protein Ure2 Enter Vertebrate Cells by Specific Endocytotic Pathways and Induce Apoptosis
title Amyloid-Like Aggregates of the Yeast Prion Protein Ure2 Enter Vertebrate Cells by Specific Endocytotic Pathways and Induce Apoptosis
title_full Amyloid-Like Aggregates of the Yeast Prion Protein Ure2 Enter Vertebrate Cells by Specific Endocytotic Pathways and Induce Apoptosis
title_fullStr Amyloid-Like Aggregates of the Yeast Prion Protein Ure2 Enter Vertebrate Cells by Specific Endocytotic Pathways and Induce Apoptosis
title_full_unstemmed Amyloid-Like Aggregates of the Yeast Prion Protein Ure2 Enter Vertebrate Cells by Specific Endocytotic Pathways and Induce Apoptosis
title_short Amyloid-Like Aggregates of the Yeast Prion Protein Ure2 Enter Vertebrate Cells by Specific Endocytotic Pathways and Induce Apoptosis
title_sort amyloid-like aggregates of the yeast prion protein ure2 enter vertebrate cells by specific endocytotic pathways and induce apoptosis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2932714/
https://www.ncbi.nlm.nih.gov/pubmed/20824085
http://dx.doi.org/10.1371/journal.pone.0012529
work_keys_str_mv AT zhangchen amyloidlikeaggregatesoftheyeastprionproteinure2entervertebratecellsbyspecificendocytoticpathwaysandinduceapoptosis
AT jacksonantonyp amyloidlikeaggregatesoftheyeastprionproteinure2entervertebratecellsbyspecificendocytoticpathwaysandinduceapoptosis
AT zhangzairong amyloidlikeaggregatesoftheyeastprionproteinure2entervertebratecellsbyspecificendocytoticpathwaysandinduceapoptosis
AT hanyan amyloidlikeaggregatesoftheyeastprionproteinure2entervertebratecellsbyspecificendocytoticpathwaysandinduceapoptosis
AT yushun amyloidlikeaggregatesoftheyeastprionproteinure2entervertebratecellsbyspecificendocytoticpathwaysandinduceapoptosis
AT herongqiao amyloidlikeaggregatesoftheyeastprionproteinure2entervertebratecellsbyspecificendocytoticpathwaysandinduceapoptosis
AT perrettsarah amyloidlikeaggregatesoftheyeastprionproteinure2entervertebratecellsbyspecificendocytoticpathwaysandinduceapoptosis