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Influence of Subcellular Localization and Functional State on Protein Turnover

Protein homeostasis is an equilibrium of paramount importance that maintains cellular performance by preserving an efficient proteome. This equilibrium avoids the accumulation of potentially toxic proteins, which could lead to cellular stress and death. While the regulators of proteostasis are the m...

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Autores principales: Yousefi, Roya, Jevdokimenko, Kristina, Kluever, Verena, Pacheu-Grau, David, Fornasiero, Eugenio F.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8306977/
https://www.ncbi.nlm.nih.gov/pubmed/34359917
http://dx.doi.org/10.3390/cells10071747
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author Yousefi, Roya
Jevdokimenko, Kristina
Kluever, Verena
Pacheu-Grau, David
Fornasiero, Eugenio F.
author_facet Yousefi, Roya
Jevdokimenko, Kristina
Kluever, Verena
Pacheu-Grau, David
Fornasiero, Eugenio F.
author_sort Yousefi, Roya
collection PubMed
description Protein homeostasis is an equilibrium of paramount importance that maintains cellular performance by preserving an efficient proteome. This equilibrium avoids the accumulation of potentially toxic proteins, which could lead to cellular stress and death. While the regulators of proteostasis are the machineries controlling protein production, folding and degradation, several other factors can influence this process. Here, we have considered two factors influencing protein turnover: the subcellular localization of a protein and its functional state. For this purpose, we used an imaging approach based on the pulse-labeling of 17 representative SNAP-tag constructs for measuring protein lifetimes. With this approach, we obtained precise measurements of protein turnover rates in several subcellular compartments. We also tested a selection of mutants modulating the function of three extensively studied proteins, the Ca(2+) sensor calmodulin, the small GTPase Rab5a and the brain creatine kinase (CKB). Finally, we followed up on the increased lifetime observed for the constitutively active Rab5a (Q79L), and we found that its stabilization correlates with enlarged endosomes and increased interaction with membranes. Overall, our data reveal that both changes in protein localization and functional state are key modulators of protein turnover, and protein lifetime fluctuations can be considered to infer changes in cellular behavior.
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spelling pubmed-83069772021-07-25 Influence of Subcellular Localization and Functional State on Protein Turnover Yousefi, Roya Jevdokimenko, Kristina Kluever, Verena Pacheu-Grau, David Fornasiero, Eugenio F. Cells Article Protein homeostasis is an equilibrium of paramount importance that maintains cellular performance by preserving an efficient proteome. This equilibrium avoids the accumulation of potentially toxic proteins, which could lead to cellular stress and death. While the regulators of proteostasis are the machineries controlling protein production, folding and degradation, several other factors can influence this process. Here, we have considered two factors influencing protein turnover: the subcellular localization of a protein and its functional state. For this purpose, we used an imaging approach based on the pulse-labeling of 17 representative SNAP-tag constructs for measuring protein lifetimes. With this approach, we obtained precise measurements of protein turnover rates in several subcellular compartments. We also tested a selection of mutants modulating the function of three extensively studied proteins, the Ca(2+) sensor calmodulin, the small GTPase Rab5a and the brain creatine kinase (CKB). Finally, we followed up on the increased lifetime observed for the constitutively active Rab5a (Q79L), and we found that its stabilization correlates with enlarged endosomes and increased interaction with membranes. Overall, our data reveal that both changes in protein localization and functional state are key modulators of protein turnover, and protein lifetime fluctuations can be considered to infer changes in cellular behavior. MDPI 2021-07-10 /pmc/articles/PMC8306977/ /pubmed/34359917 http://dx.doi.org/10.3390/cells10071747 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/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 (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Yousefi, Roya
Jevdokimenko, Kristina
Kluever, Verena
Pacheu-Grau, David
Fornasiero, Eugenio F.
Influence of Subcellular Localization and Functional State on Protein Turnover
title Influence of Subcellular Localization and Functional State on Protein Turnover
title_full Influence of Subcellular Localization and Functional State on Protein Turnover
title_fullStr Influence of Subcellular Localization and Functional State on Protein Turnover
title_full_unstemmed Influence of Subcellular Localization and Functional State on Protein Turnover
title_short Influence of Subcellular Localization and Functional State on Protein Turnover
title_sort influence of subcellular localization and functional state on protein turnover
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8306977/
https://www.ncbi.nlm.nih.gov/pubmed/34359917
http://dx.doi.org/10.3390/cells10071747
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