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p53-Dependent subcellular proteome localization following DNA damage

The nucleolus is involved in regulating several aspects of stress responses and cell cycle arrest through the tumor suppressor p53. Under normal conditions, p53 is a short-lived protein that is present in cells at a barely detectable level. Upon exposure of cells to various forms of exogenous stress...

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Autores principales: Boisvert, François-Michel, Lamond, Angus I
Formato: Online Artículo Texto
Lenguaje:English
Publicado: WILEY-VCH Verlag 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3229981/
https://www.ncbi.nlm.nih.gov/pubmed/21080495
http://dx.doi.org/10.1002/pmic.201000213
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author Boisvert, François-Michel
Lamond, Angus I
author_facet Boisvert, François-Michel
Lamond, Angus I
author_sort Boisvert, François-Michel
collection PubMed
description The nucleolus is involved in regulating several aspects of stress responses and cell cycle arrest through the tumor suppressor p53. Under normal conditions, p53 is a short-lived protein that is present in cells at a barely detectable level. Upon exposure of cells to various forms of exogenous stress, such as DNA damage, there is a stabilization of p53 which is then responsible for an ensuing cascade of events. To further investigate the effect of p53 activation, we used a MS-based proteomics method to provide an unbiased, quantitative and high-throughput approach for measuring the subcellular distribution of the proteome that is dependent on p53. The spatial proteomics method analyses a whole cell extract created by recombining differentially labeled subcellular fractions derived from cells in which proteins have been mass labeled with heavy isotopes [Boisvert, F.-M., Lam, Y. W., Lamont, D., Lamond, A. I., Mol. Cell. Proteomics 2010, 9, 457–470]. This was used here to measure the relative distribution between cytoplasm, nucleus and nucleolus of around 2000 proteins in HCT116 cells that are either expressing wild-type p53 or null for p53. Spatial proteomics also facilitates a proteome-wide comparison of changes in protein localization in response to a wide range of physiological and experimental perturbations. We used this method to study differences in protein localization in HCT116 cells either with or without p53, and studied the differences in cellular response to DNA damage following treatment of HCT116 cells with etoposide in both p53 wild-type and null genetic backgrounds.
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spelling pubmed-32299812011-12-05 p53-Dependent subcellular proteome localization following DNA damage Boisvert, François-Michel Lamond, Angus I Proteomics Research Article The nucleolus is involved in regulating several aspects of stress responses and cell cycle arrest through the tumor suppressor p53. Under normal conditions, p53 is a short-lived protein that is present in cells at a barely detectable level. Upon exposure of cells to various forms of exogenous stress, such as DNA damage, there is a stabilization of p53 which is then responsible for an ensuing cascade of events. To further investigate the effect of p53 activation, we used a MS-based proteomics method to provide an unbiased, quantitative and high-throughput approach for measuring the subcellular distribution of the proteome that is dependent on p53. The spatial proteomics method analyses a whole cell extract created by recombining differentially labeled subcellular fractions derived from cells in which proteins have been mass labeled with heavy isotopes [Boisvert, F.-M., Lam, Y. W., Lamont, D., Lamond, A. I., Mol. Cell. Proteomics 2010, 9, 457–470]. This was used here to measure the relative distribution between cytoplasm, nucleus and nucleolus of around 2000 proteins in HCT116 cells that are either expressing wild-type p53 or null for p53. Spatial proteomics also facilitates a proteome-wide comparison of changes in protein localization in response to a wide range of physiological and experimental perturbations. We used this method to study differences in protein localization in HCT116 cells either with or without p53, and studied the differences in cellular response to DNA damage following treatment of HCT116 cells with etoposide in both p53 wild-type and null genetic backgrounds. WILEY-VCH Verlag 2010-11 /pmc/articles/PMC3229981/ /pubmed/21080495 http://dx.doi.org/10.1002/pmic.201000213 Text en Copyright © 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim http://creativecommons.org/licenses/by/2.5/ Re-use of this article is permitted in accordance with the Creative Commons Deed, Attribution 2.5, which does not permit commercial exploitation.
spellingShingle Research Article
Boisvert, François-Michel
Lamond, Angus I
p53-Dependent subcellular proteome localization following DNA damage
title p53-Dependent subcellular proteome localization following DNA damage
title_full p53-Dependent subcellular proteome localization following DNA damage
title_fullStr p53-Dependent subcellular proteome localization following DNA damage
title_full_unstemmed p53-Dependent subcellular proteome localization following DNA damage
title_short p53-Dependent subcellular proteome localization following DNA damage
title_sort p53-dependent subcellular proteome localization following dna damage
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3229981/
https://www.ncbi.nlm.nih.gov/pubmed/21080495
http://dx.doi.org/10.1002/pmic.201000213
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