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Thiol stress–dependent aggregation of the glycolytic enzyme triose phosphate isomerase in yeast and human cells

The eukaryotic cytosolic proteome is vulnerable to changes in proteostatic and redox balance caused by temperature, pH, oxidants, and xenobiotics. Cysteine-containing proteins are especially at risk, as the thiol side chain is subject to oxidation, adduction, and chelation by thiol-reactive compound...

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Autores principales: Ford, Amy E., Denicourt, Catherine, Morano, Kevin A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6589699/
https://www.ncbi.nlm.nih.gov/pubmed/30601716
http://dx.doi.org/10.1091/mbc.E18-10-0616
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author Ford, Amy E.
Denicourt, Catherine
Morano, Kevin A.
author_facet Ford, Amy E.
Denicourt, Catherine
Morano, Kevin A.
author_sort Ford, Amy E.
collection PubMed
description The eukaryotic cytosolic proteome is vulnerable to changes in proteostatic and redox balance caused by temperature, pH, oxidants, and xenobiotics. Cysteine-containing proteins are especially at risk, as the thiol side chain is subject to oxidation, adduction, and chelation by thiol-reactive compounds. The thiol-chelating heavy metal cadmium is a highly toxic environmental pollutant demonstrated to induce the heat shock response and recruit protein chaperones to sites of presumed protein aggregation in the budding yeast Saccharomyces cerevisiae. However, endogenous targets of cadmium toxicity responsible for these outcomes are largely unknown. Using fluorescent protein fusion to cytosolic proteins with known redox-active cysteines, we identified the yeast glycolytic enzyme triose phosphate isomerase as being aggregation-prone in response to cadmium and to glucose depletion in chronologically aging cultures. Cadmium-induced aggregation was limited to newly synthesized Tpi1 that was recruited to foci containing the disaggregase Hsp104 and the peroxiredoxin chaperone Tsa1. Misfolding of nascent Tpi1 in response to both cadmium and glucose-depletion stress required both cysteines, implying that thiol status in this protein directly influences folding. We also demonstrate that cadmium proteotoxicity is conserved between yeast and human cells, as HEK293 and HCT116 cell lines exhibit recruitment of the protein chaperone Hsp70 to visible foci. Moreover, human TPI, mutations in which cause a glycolytic deficiency syndrome, also forms aggregates in response to cadmium treatment, suggesting that this conserved enzyme is folding-labile and may be a useful endogenous model for investigating thiol-specific proteotoxicity.
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spelling pubmed-65896992019-07-10 Thiol stress–dependent aggregation of the glycolytic enzyme triose phosphate isomerase in yeast and human cells Ford, Amy E. Denicourt, Catherine Morano, Kevin A. Mol Biol Cell Articles The eukaryotic cytosolic proteome is vulnerable to changes in proteostatic and redox balance caused by temperature, pH, oxidants, and xenobiotics. Cysteine-containing proteins are especially at risk, as the thiol side chain is subject to oxidation, adduction, and chelation by thiol-reactive compounds. The thiol-chelating heavy metal cadmium is a highly toxic environmental pollutant demonstrated to induce the heat shock response and recruit protein chaperones to sites of presumed protein aggregation in the budding yeast Saccharomyces cerevisiae. However, endogenous targets of cadmium toxicity responsible for these outcomes are largely unknown. Using fluorescent protein fusion to cytosolic proteins with known redox-active cysteines, we identified the yeast glycolytic enzyme triose phosphate isomerase as being aggregation-prone in response to cadmium and to glucose depletion in chronologically aging cultures. Cadmium-induced aggregation was limited to newly synthesized Tpi1 that was recruited to foci containing the disaggregase Hsp104 and the peroxiredoxin chaperone Tsa1. Misfolding of nascent Tpi1 in response to both cadmium and glucose-depletion stress required both cysteines, implying that thiol status in this protein directly influences folding. We also demonstrate that cadmium proteotoxicity is conserved between yeast and human cells, as HEK293 and HCT116 cell lines exhibit recruitment of the protein chaperone Hsp70 to visible foci. Moreover, human TPI, mutations in which cause a glycolytic deficiency syndrome, also forms aggregates in response to cadmium treatment, suggesting that this conserved enzyme is folding-labile and may be a useful endogenous model for investigating thiol-specific proteotoxicity. The American Society for Cell Biology 2019-03-01 /pmc/articles/PMC6589699/ /pubmed/30601716 http://dx.doi.org/10.1091/mbc.E18-10-0616 Text en © 2019 Ford et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology. http://creativecommons.org/licenses/by-nc-sa/3.0 This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License.
spellingShingle Articles
Ford, Amy E.
Denicourt, Catherine
Morano, Kevin A.
Thiol stress–dependent aggregation of the glycolytic enzyme triose phosphate isomerase in yeast and human cells
title Thiol stress–dependent aggregation of the glycolytic enzyme triose phosphate isomerase in yeast and human cells
title_full Thiol stress–dependent aggregation of the glycolytic enzyme triose phosphate isomerase in yeast and human cells
title_fullStr Thiol stress–dependent aggregation of the glycolytic enzyme triose phosphate isomerase in yeast and human cells
title_full_unstemmed Thiol stress–dependent aggregation of the glycolytic enzyme triose phosphate isomerase in yeast and human cells
title_short Thiol stress–dependent aggregation of the glycolytic enzyme triose phosphate isomerase in yeast and human cells
title_sort thiol stress–dependent aggregation of the glycolytic enzyme triose phosphate isomerase in yeast and human cells
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6589699/
https://www.ncbi.nlm.nih.gov/pubmed/30601716
http://dx.doi.org/10.1091/mbc.E18-10-0616
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