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Tryptophan 32-mediated SOD1 aggregation is attenuated by pyrimidine-like compounds in living cells

Over 160 mutations in superoxide dismutase 1 (SOD1) are associated with familial amyotrophic lateral sclerosis (fALS), where the main pathological feature is deposition of SOD1 into proteinaceous cytoplasmic inclusions. We previously showed that the tryptophan residue at position 32 (W32) mediates t...

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Autores principales: Pokrishevsky, Edward, McAlary, Luke, Farrawell, Natalie E., Zhao, Beibei, Sher, Mine, Yerbury, Justin J., Cashman, Neil R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6197196/
https://www.ncbi.nlm.nih.gov/pubmed/30349065
http://dx.doi.org/10.1038/s41598-018-32835-y
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author Pokrishevsky, Edward
McAlary, Luke
Farrawell, Natalie E.
Zhao, Beibei
Sher, Mine
Yerbury, Justin J.
Cashman, Neil R.
author_facet Pokrishevsky, Edward
McAlary, Luke
Farrawell, Natalie E.
Zhao, Beibei
Sher, Mine
Yerbury, Justin J.
Cashman, Neil R.
author_sort Pokrishevsky, Edward
collection PubMed
description Over 160 mutations in superoxide dismutase 1 (SOD1) are associated with familial amyotrophic lateral sclerosis (fALS), where the main pathological feature is deposition of SOD1 into proteinaceous cytoplasmic inclusions. We previously showed that the tryptophan residue at position 32 (W32) mediates the prion-like propagation of SOD1 misfolding in cells, and that a W32S substitution blocks this phenomenon. Here, we used in vitro protein assays to demonstrate that a W32S substitution in SOD1-fALS mutants significantly diminishes their propensity to aggregate whilst paradoxically decreasing protein stability. We also show SOD1-W32S to be resistant to seeded aggregation, despite its high abundance of unfolded protein. A cell-based aggregation assay demonstrates that W32S substitution significantly mitigates inclusion formation. Furthermore, this assay reveals that W32 in SOD1 is necessary for the formation of a competent seed for aggregation under these experimental conditions. Following the observed importance of W32 for aggregation, we established that treatment of living cells with the W32-interacting 5-Fluorouridine (5-FUrd), and its FDA approved analogue 5-Fluorouracil (5-FU), substantially attenuate inclusion formation similarly to W32S substitution. Altogether, we highlight W32 as a significant contributor to SOD1 aggregation, and propose that 5-FUrd and 5-FU present promising lead drug candidates for the treatment of SOD1-associated ALS.
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spelling pubmed-61971962018-10-24 Tryptophan 32-mediated SOD1 aggregation is attenuated by pyrimidine-like compounds in living cells Pokrishevsky, Edward McAlary, Luke Farrawell, Natalie E. Zhao, Beibei Sher, Mine Yerbury, Justin J. Cashman, Neil R. Sci Rep Article Over 160 mutations in superoxide dismutase 1 (SOD1) are associated with familial amyotrophic lateral sclerosis (fALS), where the main pathological feature is deposition of SOD1 into proteinaceous cytoplasmic inclusions. We previously showed that the tryptophan residue at position 32 (W32) mediates the prion-like propagation of SOD1 misfolding in cells, and that a W32S substitution blocks this phenomenon. Here, we used in vitro protein assays to demonstrate that a W32S substitution in SOD1-fALS mutants significantly diminishes their propensity to aggregate whilst paradoxically decreasing protein stability. We also show SOD1-W32S to be resistant to seeded aggregation, despite its high abundance of unfolded protein. A cell-based aggregation assay demonstrates that W32S substitution significantly mitigates inclusion formation. Furthermore, this assay reveals that W32 in SOD1 is necessary for the formation of a competent seed for aggregation under these experimental conditions. Following the observed importance of W32 for aggregation, we established that treatment of living cells with the W32-interacting 5-Fluorouridine (5-FUrd), and its FDA approved analogue 5-Fluorouracil (5-FU), substantially attenuate inclusion formation similarly to W32S substitution. Altogether, we highlight W32 as a significant contributor to SOD1 aggregation, and propose that 5-FUrd and 5-FU present promising lead drug candidates for the treatment of SOD1-associated ALS. Nature Publishing Group UK 2018-10-22 /pmc/articles/PMC6197196/ /pubmed/30349065 http://dx.doi.org/10.1038/s41598-018-32835-y Text en © The Author(s) 2018 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Pokrishevsky, Edward
McAlary, Luke
Farrawell, Natalie E.
Zhao, Beibei
Sher, Mine
Yerbury, Justin J.
Cashman, Neil R.
Tryptophan 32-mediated SOD1 aggregation is attenuated by pyrimidine-like compounds in living cells
title Tryptophan 32-mediated SOD1 aggregation is attenuated by pyrimidine-like compounds in living cells
title_full Tryptophan 32-mediated SOD1 aggregation is attenuated by pyrimidine-like compounds in living cells
title_fullStr Tryptophan 32-mediated SOD1 aggregation is attenuated by pyrimidine-like compounds in living cells
title_full_unstemmed Tryptophan 32-mediated SOD1 aggregation is attenuated by pyrimidine-like compounds in living cells
title_short Tryptophan 32-mediated SOD1 aggregation is attenuated by pyrimidine-like compounds in living cells
title_sort tryptophan 32-mediated sod1 aggregation is attenuated by pyrimidine-like compounds in living cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6197196/
https://www.ncbi.nlm.nih.gov/pubmed/30349065
http://dx.doi.org/10.1038/s41598-018-32835-y
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