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The polyglutamine protein ataxin-3 enables normal growth under heat shock conditions in the methylotrophic yeast Pichia pastoris

The protein ataxin-3 carries a polyglutamine stretch close to the C-terminus that triggers a neurodegenerative disease in humans when its length exceeds a critical threshold. A role as a transcriptional regulator but also as a ubiquitin hydrolase has been proposed for this protein. Here, we report t...

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Autores principales: Bonanomi, Marcella, Roffia, Valentina, De Palma, Antonella, Lombardi, Alessio, Aprile, Francesco Antonio, Visentin, Cristina, Tortora, Paolo, Mauri, Pierluigi, Regonesi, Maria Elena
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5645362/
https://www.ncbi.nlm.nih.gov/pubmed/29042637
http://dx.doi.org/10.1038/s41598-017-13814-1
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author Bonanomi, Marcella
Roffia, Valentina
De Palma, Antonella
Lombardi, Alessio
Aprile, Francesco Antonio
Visentin, Cristina
Tortora, Paolo
Mauri, Pierluigi
Regonesi, Maria Elena
author_facet Bonanomi, Marcella
Roffia, Valentina
De Palma, Antonella
Lombardi, Alessio
Aprile, Francesco Antonio
Visentin, Cristina
Tortora, Paolo
Mauri, Pierluigi
Regonesi, Maria Elena
author_sort Bonanomi, Marcella
collection PubMed
description The protein ataxin-3 carries a polyglutamine stretch close to the C-terminus that triggers a neurodegenerative disease in humans when its length exceeds a critical threshold. A role as a transcriptional regulator but also as a ubiquitin hydrolase has been proposed for this protein. Here, we report that, when expressed in the yeast Pichia pastoris, full-length ataxin-3 enabled almost normal growth at 37 °C, well above the physiological optimum of 30 °C. The N-terminal Josephin domain (JD) was also effective but significantly less, whereas catalytically inactive JD was completely ineffective. Based on MudPIT proteomic analysis, we observed that the strain expressing full-length, functional ataxin-3 displayed persistent upregulation of enzymes involved in mitochondrial energy metabolism during growth at 37 °C compared with the strain transformed with the empty vector. Concurrently, in the transformed strain intracellular ATP levels at 37 °C were even higher than normal ones at 30 °C. Elevated ATP was also paralleled by upregulation of enzymes involved in both protein biosynthesis and biosynthetic pathways, as well as of several stress-induced proteins. A similar pattern was observed when comparing a strain expressing JD with another expressing its catalytically inactive counterpart. We suggest that such effects mostly result from mechanisms of transcriptional regulation.
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spelling pubmed-56453622017-10-26 The polyglutamine protein ataxin-3 enables normal growth under heat shock conditions in the methylotrophic yeast Pichia pastoris Bonanomi, Marcella Roffia, Valentina De Palma, Antonella Lombardi, Alessio Aprile, Francesco Antonio Visentin, Cristina Tortora, Paolo Mauri, Pierluigi Regonesi, Maria Elena Sci Rep Article The protein ataxin-3 carries a polyglutamine stretch close to the C-terminus that triggers a neurodegenerative disease in humans when its length exceeds a critical threshold. A role as a transcriptional regulator but also as a ubiquitin hydrolase has been proposed for this protein. Here, we report that, when expressed in the yeast Pichia pastoris, full-length ataxin-3 enabled almost normal growth at 37 °C, well above the physiological optimum of 30 °C. The N-terminal Josephin domain (JD) was also effective but significantly less, whereas catalytically inactive JD was completely ineffective. Based on MudPIT proteomic analysis, we observed that the strain expressing full-length, functional ataxin-3 displayed persistent upregulation of enzymes involved in mitochondrial energy metabolism during growth at 37 °C compared with the strain transformed with the empty vector. Concurrently, in the transformed strain intracellular ATP levels at 37 °C were even higher than normal ones at 30 °C. Elevated ATP was also paralleled by upregulation of enzymes involved in both protein biosynthesis and biosynthetic pathways, as well as of several stress-induced proteins. A similar pattern was observed when comparing a strain expressing JD with another expressing its catalytically inactive counterpart. We suggest that such effects mostly result from mechanisms of transcriptional regulation. Nature Publishing Group UK 2017-10-17 /pmc/articles/PMC5645362/ /pubmed/29042637 http://dx.doi.org/10.1038/s41598-017-13814-1 Text en © The Author(s) 2017 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
Bonanomi, Marcella
Roffia, Valentina
De Palma, Antonella
Lombardi, Alessio
Aprile, Francesco Antonio
Visentin, Cristina
Tortora, Paolo
Mauri, Pierluigi
Regonesi, Maria Elena
The polyglutamine protein ataxin-3 enables normal growth under heat shock conditions in the methylotrophic yeast Pichia pastoris
title The polyglutamine protein ataxin-3 enables normal growth under heat shock conditions in the methylotrophic yeast Pichia pastoris
title_full The polyglutamine protein ataxin-3 enables normal growth under heat shock conditions in the methylotrophic yeast Pichia pastoris
title_fullStr The polyglutamine protein ataxin-3 enables normal growth under heat shock conditions in the methylotrophic yeast Pichia pastoris
title_full_unstemmed The polyglutamine protein ataxin-3 enables normal growth under heat shock conditions in the methylotrophic yeast Pichia pastoris
title_short The polyglutamine protein ataxin-3 enables normal growth under heat shock conditions in the methylotrophic yeast Pichia pastoris
title_sort polyglutamine protein ataxin-3 enables normal growth under heat shock conditions in the methylotrophic yeast pichia pastoris
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5645362/
https://www.ncbi.nlm.nih.gov/pubmed/29042637
http://dx.doi.org/10.1038/s41598-017-13814-1
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