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DNA damage-induced ubiquitylation of proteasome controls its proteolytic activity
Stability of proteins is largely controlled by post-translational covalent modifications. Among those, ubiquitylation plays a central role as it marks the proteins for proteasome-dependent degradation. Proteolytic activities of proteasomes are critical for execution of various cellular processes, in...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Impact Journals LLC
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3824523/ https://www.ncbi.nlm.nih.gov/pubmed/23907514 |
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author | Moiseeva, Tatiana N. Bottrill, Andrew Melino, Gerry Barlev, Nickolai A. |
author_facet | Moiseeva, Tatiana N. Bottrill, Andrew Melino, Gerry Barlev, Nickolai A. |
author_sort | Moiseeva, Tatiana N. |
collection | PubMed |
description | Stability of proteins is largely controlled by post-translational covalent modifications. Among those, ubiquitylation plays a central role as it marks the proteins for proteasome-dependent degradation. Proteolytic activities of proteasomes are critical for execution of various cellular processes, including DNA damage signaling and repair. However, very little is known about the regulation of proteasomal activity in cells during genotoxic stress. Here we investigated post-translational modifications of the 20S proteasomal subunits upon DNA damage induced by doxorubicin. Using mass-spectrometry, we found novel sites of phosphorylation and ubiquitylation in multiple proteasome subunits upon doxorubicin treatment. Ectopic co-expression of proteasome subunits and tagged ubiquitin confirmed the presence of ubiquitylated forms of PSMA5, PSMA1, PSMA3 and PSMB5 in cells. Moreover, we demonstrated that ubiquitylation in vitro inhibited chymotrypsin-like and caspase-like activities of proteasomes. In vivo, doxorubicin increased the activity of proteasomes, paralleling with attenuation of the overall level of proteasome ubiquitylation. Collectively, our results suggest a novel mechanism whereby the proteolytic activities of proteasomes are dynamically regulated by ubiquitylation upon DNA damage. |
format | Online Article Text |
id | pubmed-3824523 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Impact Journals LLC |
record_format | MEDLINE/PubMed |
spelling | pubmed-38245232013-11-22 DNA damage-induced ubiquitylation of proteasome controls its proteolytic activity Moiseeva, Tatiana N. Bottrill, Andrew Melino, Gerry Barlev, Nickolai A. Oncotarget Research Paper Stability of proteins is largely controlled by post-translational covalent modifications. Among those, ubiquitylation plays a central role as it marks the proteins for proteasome-dependent degradation. Proteolytic activities of proteasomes are critical for execution of various cellular processes, including DNA damage signaling and repair. However, very little is known about the regulation of proteasomal activity in cells during genotoxic stress. Here we investigated post-translational modifications of the 20S proteasomal subunits upon DNA damage induced by doxorubicin. Using mass-spectrometry, we found novel sites of phosphorylation and ubiquitylation in multiple proteasome subunits upon doxorubicin treatment. Ectopic co-expression of proteasome subunits and tagged ubiquitin confirmed the presence of ubiquitylated forms of PSMA5, PSMA1, PSMA3 and PSMB5 in cells. Moreover, we demonstrated that ubiquitylation in vitro inhibited chymotrypsin-like and caspase-like activities of proteasomes. In vivo, doxorubicin increased the activity of proteasomes, paralleling with attenuation of the overall level of proteasome ubiquitylation. Collectively, our results suggest a novel mechanism whereby the proteolytic activities of proteasomes are dynamically regulated by ubiquitylation upon DNA damage. Impact Journals LLC 2013-06-29 /pmc/articles/PMC3824523/ /pubmed/23907514 Text en Copyright: © 2013 Moiseeva et al. http://creativecommons.org/licenses/by/2.5/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited |
spellingShingle | Research Paper Moiseeva, Tatiana N. Bottrill, Andrew Melino, Gerry Barlev, Nickolai A. DNA damage-induced ubiquitylation of proteasome controls its proteolytic activity |
title | DNA damage-induced ubiquitylation of proteasome controls its proteolytic activity |
title_full | DNA damage-induced ubiquitylation of proteasome controls its proteolytic activity |
title_fullStr | DNA damage-induced ubiquitylation of proteasome controls its proteolytic activity |
title_full_unstemmed | DNA damage-induced ubiquitylation of proteasome controls its proteolytic activity |
title_short | DNA damage-induced ubiquitylation of proteasome controls its proteolytic activity |
title_sort | dna damage-induced ubiquitylation of proteasome controls its proteolytic activity |
topic | Research Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3824523/ https://www.ncbi.nlm.nih.gov/pubmed/23907514 |
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