Lipid rafts and clathrin cooperate in the internalization of PrP in epithelial FRT cells.
<h4>Background</h4>The cellular prion protein (PrP(C)) plays a key role in the pathogenesis of Transmissible Spongiform Encephalopathies in which the protein undergoes post-translational conversion to the infectious form (PrP(Sc)). Although endocytosis appears to be required for this con...
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Public Library of Science (PLoS)
2009-06-01
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| Series: | PLoS ONE |
| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0005829&type=printable |
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| author | Daniela Sarnataro Anna Caputo Philippe Casanova Claudia Puri Simona Paladino Simona S Tivodar Vincenza Campana Carlo Tacchetti Chiara Zurzolo |
| author_facet | Daniela Sarnataro Anna Caputo Philippe Casanova Claudia Puri Simona Paladino Simona S Tivodar Vincenza Campana Carlo Tacchetti Chiara Zurzolo |
| author_sort | Daniela Sarnataro |
| collection | DOAJ |
| description | <h4>Background</h4>The cellular prion protein (PrP(C)) plays a key role in the pathogenesis of Transmissible Spongiform Encephalopathies in which the protein undergoes post-translational conversion to the infectious form (PrP(Sc)). Although endocytosis appears to be required for this conversion, the mechanism of PrP(C) internalization is still debated, as caveolae/raft- and clathrin-dependent processes have all been reported to be involved.<h4>Methodology/principal findings</h4>We have investigated the mechanism of PrP(C) endocytosis in Fischer Rat Thyroid (FRT) cells, which lack caveolin-1 (cav-1) and caveolae, and in FRT/cav-1 cells which form functional caveolae. We show that PrP(C) internalization requires activated Cdc-42 and is sensitive to cholesterol depletion but not to cav-1 expression suggesting a role for rafts but not for caveolae in PrP(C) endocytosis. PrP(C) internalization is also affected by knock down of clathrin and by the expression of dominant negative Eps15 and Dynamin 2 mutants, indicating the involvement of a clathrin-dependent pathway. Notably, PrP(C) co-immunoprecipitates with clathrin and remains associated with detergent-insoluble microdomains during internalization thus indicating that PrP(C) can enter the cell via multiple pathways and that rafts and clathrin cooperate in its internalization.<h4>Conclusions/significance</h4>These findings are of particular interest if we consider that the internalization route/s undertaken by PrP(C) can be crucial for the ability of different prion strains to infect and to replicate in different cell lines. |
| format | Article |
| id | doaj-art-d4e5b6c14de444668997aa87a5840bc7 |
| institution | OA Journals |
| issn | 1932-6203 |
| language | English |
| publishDate | 2009-06-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-d4e5b6c14de444668997aa87a5840bc72025-08-20T02:00:52ZengPublic Library of Science (PLoS)PLoS ONE1932-62032009-06-0146e582910.1371/journal.pone.0005829Lipid rafts and clathrin cooperate in the internalization of PrP in epithelial FRT cells.Daniela SarnataroAnna CaputoPhilippe CasanovaClaudia PuriSimona PaladinoSimona S TivodarVincenza CampanaCarlo TacchettiChiara Zurzolo<h4>Background</h4>The cellular prion protein (PrP(C)) plays a key role in the pathogenesis of Transmissible Spongiform Encephalopathies in which the protein undergoes post-translational conversion to the infectious form (PrP(Sc)). Although endocytosis appears to be required for this conversion, the mechanism of PrP(C) internalization is still debated, as caveolae/raft- and clathrin-dependent processes have all been reported to be involved.<h4>Methodology/principal findings</h4>We have investigated the mechanism of PrP(C) endocytosis in Fischer Rat Thyroid (FRT) cells, which lack caveolin-1 (cav-1) and caveolae, and in FRT/cav-1 cells which form functional caveolae. We show that PrP(C) internalization requires activated Cdc-42 and is sensitive to cholesterol depletion but not to cav-1 expression suggesting a role for rafts but not for caveolae in PrP(C) endocytosis. PrP(C) internalization is also affected by knock down of clathrin and by the expression of dominant negative Eps15 and Dynamin 2 mutants, indicating the involvement of a clathrin-dependent pathway. Notably, PrP(C) co-immunoprecipitates with clathrin and remains associated with detergent-insoluble microdomains during internalization thus indicating that PrP(C) can enter the cell via multiple pathways and that rafts and clathrin cooperate in its internalization.<h4>Conclusions/significance</h4>These findings are of particular interest if we consider that the internalization route/s undertaken by PrP(C) can be crucial for the ability of different prion strains to infect and to replicate in different cell lines.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0005829&type=printable |
| spellingShingle | Daniela Sarnataro Anna Caputo Philippe Casanova Claudia Puri Simona Paladino Simona S Tivodar Vincenza Campana Carlo Tacchetti Chiara Zurzolo Lipid rafts and clathrin cooperate in the internalization of PrP in epithelial FRT cells. PLoS ONE |
| title | Lipid rafts and clathrin cooperate in the internalization of PrP in epithelial FRT cells. |
| title_full | Lipid rafts and clathrin cooperate in the internalization of PrP in epithelial FRT cells. |
| title_fullStr | Lipid rafts and clathrin cooperate in the internalization of PrP in epithelial FRT cells. |
| title_full_unstemmed | Lipid rafts and clathrin cooperate in the internalization of PrP in epithelial FRT cells. |
| title_short | Lipid rafts and clathrin cooperate in the internalization of PrP in epithelial FRT cells. |
| title_sort | lipid rafts and clathrin cooperate in the internalization of prp in epithelial frt cells |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0005829&type=printable |
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