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Critical Significance of the Region between Helix 1 and 2 for Efficient Dominant-Negative Inhibition by Conversion-Incompetent Prion Protein

Prion diseases are fatal infectious neurodegenerative disorders in man and animals associated with the accumulation of the pathogenic isoform PrP(Sc) of the host-encoded prion protein (PrP(c)). A profound conformational change of PrP(c) underlies formation of PrP(Sc) and prion propagation involves c...

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Autores principales: Taguchi, Yuzuru, Mistica, Arla M. A., Kitamoto, Tetsuyuki, Schätzl, Hermann M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3694865/
https://www.ncbi.nlm.nih.gov/pubmed/23825952
http://dx.doi.org/10.1371/journal.ppat.1003466
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author Taguchi, Yuzuru
Mistica, Arla M. A.
Kitamoto, Tetsuyuki
Schätzl, Hermann M.
author_facet Taguchi, Yuzuru
Mistica, Arla M. A.
Kitamoto, Tetsuyuki
Schätzl, Hermann M.
author_sort Taguchi, Yuzuru
collection PubMed
description Prion diseases are fatal infectious neurodegenerative disorders in man and animals associated with the accumulation of the pathogenic isoform PrP(Sc) of the host-encoded prion protein (PrP(c)). A profound conformational change of PrP(c) underlies formation of PrP(Sc) and prion propagation involves conversion of PrP(c) substrate by direct interaction with PrP(Sc) template. Identifying the interfaces and modalities of inter-molecular interactions of PrPs will highly advance our understanding of prion propagation in particular and of prion-like mechanisms in general. To identify the region critical for inter-molecular interactions of PrP, we exploited here dominant-negative inhibition (DNI) effects of conversion-incompetent, internally-deleted PrP (ΔPrP) on co-expressed conversion-competent PrP. We created a series of ΔPrPs with different lengths of deletions in the region between first and second α-helix (H1∼H2) which was recently postulated to be of importance in prion species barrier and PrP fibril formation. As previously reported, ΔPrPs uniformly exhibited aberrant properties including detergent insolubility, limited protease digestion resistance, high-mannose type N-linked glycans, and intracellular localization. Although formerly controversial, we demonstrate here that ΔPrPs have a GPI anchor attached. Surprisingly, despite very similar biochemical and cell-biological properties, DNI efficiencies of ΔPrPs varied significantly, dependant on location and inversely correlated with the size of deletion. This data demonstrates that H1∼H2 and the region C-terminal to it are critically important for efficient DNI. It also suggests that this region is involved in PrP-PrP interaction and conversion of PrP(C) into PrP(Sc). To reconcile the paradox of how an intracellular PrP can exert DNI, we demonstrate that ΔPrPs are subject to both proteasomal and lysosomal/autophagic degradation pathways. Using autophagy pathways ΔPrPs obtain access to the locale of prion conversion and PrP(Sc) recycling and can exert DNI there. This shows that the intracellular trafficking of PrPs is more complex than previously anticipated.
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spelling pubmed-36948652013-07-03 Critical Significance of the Region between Helix 1 and 2 for Efficient Dominant-Negative Inhibition by Conversion-Incompetent Prion Protein Taguchi, Yuzuru Mistica, Arla M. A. Kitamoto, Tetsuyuki Schätzl, Hermann M. PLoS Pathog Research Article Prion diseases are fatal infectious neurodegenerative disorders in man and animals associated with the accumulation of the pathogenic isoform PrP(Sc) of the host-encoded prion protein (PrP(c)). A profound conformational change of PrP(c) underlies formation of PrP(Sc) and prion propagation involves conversion of PrP(c) substrate by direct interaction with PrP(Sc) template. Identifying the interfaces and modalities of inter-molecular interactions of PrPs will highly advance our understanding of prion propagation in particular and of prion-like mechanisms in general. To identify the region critical for inter-molecular interactions of PrP, we exploited here dominant-negative inhibition (DNI) effects of conversion-incompetent, internally-deleted PrP (ΔPrP) on co-expressed conversion-competent PrP. We created a series of ΔPrPs with different lengths of deletions in the region between first and second α-helix (H1∼H2) which was recently postulated to be of importance in prion species barrier and PrP fibril formation. As previously reported, ΔPrPs uniformly exhibited aberrant properties including detergent insolubility, limited protease digestion resistance, high-mannose type N-linked glycans, and intracellular localization. Although formerly controversial, we demonstrate here that ΔPrPs have a GPI anchor attached. Surprisingly, despite very similar biochemical and cell-biological properties, DNI efficiencies of ΔPrPs varied significantly, dependant on location and inversely correlated with the size of deletion. This data demonstrates that H1∼H2 and the region C-terminal to it are critically important for efficient DNI. It also suggests that this region is involved in PrP-PrP interaction and conversion of PrP(C) into PrP(Sc). To reconcile the paradox of how an intracellular PrP can exert DNI, we demonstrate that ΔPrPs are subject to both proteasomal and lysosomal/autophagic degradation pathways. Using autophagy pathways ΔPrPs obtain access to the locale of prion conversion and PrP(Sc) recycling and can exert DNI there. This shows that the intracellular trafficking of PrPs is more complex than previously anticipated. Public Library of Science 2013-06-27 /pmc/articles/PMC3694865/ /pubmed/23825952 http://dx.doi.org/10.1371/journal.ppat.1003466 Text en © 2013 Taguchi 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
Taguchi, Yuzuru
Mistica, Arla M. A.
Kitamoto, Tetsuyuki
Schätzl, Hermann M.
Critical Significance of the Region between Helix 1 and 2 for Efficient Dominant-Negative Inhibition by Conversion-Incompetent Prion Protein
title Critical Significance of the Region between Helix 1 and 2 for Efficient Dominant-Negative Inhibition by Conversion-Incompetent Prion Protein
title_full Critical Significance of the Region between Helix 1 and 2 for Efficient Dominant-Negative Inhibition by Conversion-Incompetent Prion Protein
title_fullStr Critical Significance of the Region between Helix 1 and 2 for Efficient Dominant-Negative Inhibition by Conversion-Incompetent Prion Protein
title_full_unstemmed Critical Significance of the Region between Helix 1 and 2 for Efficient Dominant-Negative Inhibition by Conversion-Incompetent Prion Protein
title_short Critical Significance of the Region between Helix 1 and 2 for Efficient Dominant-Negative Inhibition by Conversion-Incompetent Prion Protein
title_sort critical significance of the region between helix 1 and 2 for efficient dominant-negative inhibition by conversion-incompetent prion protein
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3694865/
https://www.ncbi.nlm.nih.gov/pubmed/23825952
http://dx.doi.org/10.1371/journal.ppat.1003466
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