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The N-Terminal Region of Yeast Protein Phosphatase Ppz1 Is a Determinant for Its Toxicity

The Ppz enzymes are Ser/Thr protein phosphatases present only in fungi that are characterized by a highly conserved C-terminal catalytic region, related to PP1c phosphatases, and a more divergent N-terminal extension. In Saccharomyces cerevisiae, Ppz phosphatases are encoded by two paralog genes, PP...

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Autores principales: Calafí, Carlos, López-Malo, María, Albacar, Marcel, Casamayor, Antonio, Ariño, Joaquín
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7590015/
https://www.ncbi.nlm.nih.gov/pubmed/33086699
http://dx.doi.org/10.3390/ijms21207733
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author Calafí, Carlos
López-Malo, María
Albacar, Marcel
Casamayor, Antonio
Ariño, Joaquín
author_facet Calafí, Carlos
López-Malo, María
Albacar, Marcel
Casamayor, Antonio
Ariño, Joaquín
author_sort Calafí, Carlos
collection PubMed
description The Ppz enzymes are Ser/Thr protein phosphatases present only in fungi that are characterized by a highly conserved C-terminal catalytic region, related to PP1c phosphatases, and a more divergent N-terminal extension. In Saccharomyces cerevisiae, Ppz phosphatases are encoded by two paralog genes, PPZ1 and PPZ2. Ppz1 is the most toxic protein when overexpressed in budding yeast, halting cell proliferation, and this effect requires its phosphatase activity. We show here that, in spite of their conserved catalytic domain, Ppz2 was not toxic when tested under the same conditions as Ppz1, albeit Ppz2 levels were somewhat lower. Remarkably, a hybrid protein composed of the N-terminal extension of Ppz1 and the catalytic domain of Ppz2 was as toxic as Ppz1, even if its expression level was comparable to that of Ppz2. Similar amounts of yeast PP1c (Glc7) produced an intermediate effect on growth. Mutation of the Ppz1 myristoylable Gly2 to Ala avoided the localization of the phosphatase at the cell periphery but only slightly attenuated its toxicity. Therefore, the N-terminal extension of Ppz1 plays a key role in defining Ppz1 toxicity. This region is predicted to be intrinsically disordered and contains several putative folding-upon-binding regions which are absent in Ppz2 and might be relevant for toxicity.
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spelling pubmed-75900152020-10-29 The N-Terminal Region of Yeast Protein Phosphatase Ppz1 Is a Determinant for Its Toxicity Calafí, Carlos López-Malo, María Albacar, Marcel Casamayor, Antonio Ariño, Joaquín Int J Mol Sci Article The Ppz enzymes are Ser/Thr protein phosphatases present only in fungi that are characterized by a highly conserved C-terminal catalytic region, related to PP1c phosphatases, and a more divergent N-terminal extension. In Saccharomyces cerevisiae, Ppz phosphatases are encoded by two paralog genes, PPZ1 and PPZ2. Ppz1 is the most toxic protein when overexpressed in budding yeast, halting cell proliferation, and this effect requires its phosphatase activity. We show here that, in spite of their conserved catalytic domain, Ppz2 was not toxic when tested under the same conditions as Ppz1, albeit Ppz2 levels were somewhat lower. Remarkably, a hybrid protein composed of the N-terminal extension of Ppz1 and the catalytic domain of Ppz2 was as toxic as Ppz1, even if its expression level was comparable to that of Ppz2. Similar amounts of yeast PP1c (Glc7) produced an intermediate effect on growth. Mutation of the Ppz1 myristoylable Gly2 to Ala avoided the localization of the phosphatase at the cell periphery but only slightly attenuated its toxicity. Therefore, the N-terminal extension of Ppz1 plays a key role in defining Ppz1 toxicity. This region is predicted to be intrinsically disordered and contains several putative folding-upon-binding regions which are absent in Ppz2 and might be relevant for toxicity. MDPI 2020-10-19 /pmc/articles/PMC7590015/ /pubmed/33086699 http://dx.doi.org/10.3390/ijms21207733 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Calafí, Carlos
López-Malo, María
Albacar, Marcel
Casamayor, Antonio
Ariño, Joaquín
The N-Terminal Region of Yeast Protein Phosphatase Ppz1 Is a Determinant for Its Toxicity
title The N-Terminal Region of Yeast Protein Phosphatase Ppz1 Is a Determinant for Its Toxicity
title_full The N-Terminal Region of Yeast Protein Phosphatase Ppz1 Is a Determinant for Its Toxicity
title_fullStr The N-Terminal Region of Yeast Protein Phosphatase Ppz1 Is a Determinant for Its Toxicity
title_full_unstemmed The N-Terminal Region of Yeast Protein Phosphatase Ppz1 Is a Determinant for Its Toxicity
title_short The N-Terminal Region of Yeast Protein Phosphatase Ppz1 Is a Determinant for Its Toxicity
title_sort n-terminal region of yeast protein phosphatase ppz1 is a determinant for its toxicity
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7590015/
https://www.ncbi.nlm.nih.gov/pubmed/33086699
http://dx.doi.org/10.3390/ijms21207733
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