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Identification of deubiquitinase targets of isothiocyanates using SILAC-assisted quantitative mass spectrometry

Cruciferous vegetables such as broccoli and kale have well documented chemopreventative and anticancer effects that are attributed to the presence of isothiocyanates (ITCs). ITCs modulate the levels of many oncogenic proteins, but the molecular mechanisms of ITC action are not understood. We previou...

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Autores principales: Lawson, Ann P., Bak, Daniel W., Shannon, D. Alexander, Long, Marcus J.C., Vijaykumar, Tushara, Yu, Runhan, Oualid, Farid El, Weerapana, Eranthie, Hedstrom, Lizbeth
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Impact Journals LLC 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5584250/
https://www.ncbi.nlm.nih.gov/pubmed/28881649
http://dx.doi.org/10.18632/oncotarget.17261
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author Lawson, Ann P.
Bak, Daniel W.
Shannon, D. Alexander
Long, Marcus J.C.
Vijaykumar, Tushara
Yu, Runhan
Oualid, Farid El
Weerapana, Eranthie
Hedstrom, Lizbeth
author_facet Lawson, Ann P.
Bak, Daniel W.
Shannon, D. Alexander
Long, Marcus J.C.
Vijaykumar, Tushara
Yu, Runhan
Oualid, Farid El
Weerapana, Eranthie
Hedstrom, Lizbeth
author_sort Lawson, Ann P.
collection PubMed
description Cruciferous vegetables such as broccoli and kale have well documented chemopreventative and anticancer effects that are attributed to the presence of isothiocyanates (ITCs). ITCs modulate the levels of many oncogenic proteins, but the molecular mechanisms of ITC action are not understood. We previously reported that phenethyl isothiocyanate (PEITC) inhibits two deubiquitinases (DUBs), USP9x and UCH37. DUBs regulate many cellular processes and DUB dysregulation is linked to the pathogenesis of human diseases including cancer, neurodegeneration, and inflammation. Using SILAC assisted quantitative mass spectrometry, here we identify 9 new PEITC-DUB targets: USP1, USP3, USP10, USP11, USP16, USP22, USP40, USP48 and VCPIP1. Seven of these PEITC-sensitive DUBs have well-recognized roles in DNA repair or chromatin remodeling. PEITC both inhibits USP1 and increases its ubiquitination and degradation, thus decreasing USP1 activity by two mechanisms. The loss of USP1 activity increases the level of mono-ubiquitinated DNA clamp PCNA, impairing DNA repair. Both the inhibition/degradation of USP1 and the increase in mono-ubiquitinated PCNA are new activities for PEITC that can explain the previously recognized ability of ITCs to enhance cancer cell sensitivity to cisplatin treatment. Our work also demonstrates that PEITC reduces the mono-ubiquityl histones H2A and H2B. Understanding the mechanism of action of ITCs should facilitate their use as therapeutic agents.
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spelling pubmed-55842502017-09-06 Identification of deubiquitinase targets of isothiocyanates using SILAC-assisted quantitative mass spectrometry Lawson, Ann P. Bak, Daniel W. Shannon, D. Alexander Long, Marcus J.C. Vijaykumar, Tushara Yu, Runhan Oualid, Farid El Weerapana, Eranthie Hedstrom, Lizbeth Oncotarget Research Paper Cruciferous vegetables such as broccoli and kale have well documented chemopreventative and anticancer effects that are attributed to the presence of isothiocyanates (ITCs). ITCs modulate the levels of many oncogenic proteins, but the molecular mechanisms of ITC action are not understood. We previously reported that phenethyl isothiocyanate (PEITC) inhibits two deubiquitinases (DUBs), USP9x and UCH37. DUBs regulate many cellular processes and DUB dysregulation is linked to the pathogenesis of human diseases including cancer, neurodegeneration, and inflammation. Using SILAC assisted quantitative mass spectrometry, here we identify 9 new PEITC-DUB targets: USP1, USP3, USP10, USP11, USP16, USP22, USP40, USP48 and VCPIP1. Seven of these PEITC-sensitive DUBs have well-recognized roles in DNA repair or chromatin remodeling. PEITC both inhibits USP1 and increases its ubiquitination and degradation, thus decreasing USP1 activity by two mechanisms. The loss of USP1 activity increases the level of mono-ubiquitinated DNA clamp PCNA, impairing DNA repair. Both the inhibition/degradation of USP1 and the increase in mono-ubiquitinated PCNA are new activities for PEITC that can explain the previously recognized ability of ITCs to enhance cancer cell sensitivity to cisplatin treatment. Our work also demonstrates that PEITC reduces the mono-ubiquityl histones H2A and H2B. Understanding the mechanism of action of ITCs should facilitate their use as therapeutic agents. Impact Journals LLC 2017-04-20 /pmc/articles/PMC5584250/ /pubmed/28881649 http://dx.doi.org/10.18632/oncotarget.17261 Text en Copyright: © 2017 Lawson et al. http://creativecommons.org/licenses/by/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License 3.0 (http://creativecommons.org/licenses/by/3.0/) (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Paper
Lawson, Ann P.
Bak, Daniel W.
Shannon, D. Alexander
Long, Marcus J.C.
Vijaykumar, Tushara
Yu, Runhan
Oualid, Farid El
Weerapana, Eranthie
Hedstrom, Lizbeth
Identification of deubiquitinase targets of isothiocyanates using SILAC-assisted quantitative mass spectrometry
title Identification of deubiquitinase targets of isothiocyanates using SILAC-assisted quantitative mass spectrometry
title_full Identification of deubiquitinase targets of isothiocyanates using SILAC-assisted quantitative mass spectrometry
title_fullStr Identification of deubiquitinase targets of isothiocyanates using SILAC-assisted quantitative mass spectrometry
title_full_unstemmed Identification of deubiquitinase targets of isothiocyanates using SILAC-assisted quantitative mass spectrometry
title_short Identification of deubiquitinase targets of isothiocyanates using SILAC-assisted quantitative mass spectrometry
title_sort identification of deubiquitinase targets of isothiocyanates using silac-assisted quantitative mass spectrometry
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5584250/
https://www.ncbi.nlm.nih.gov/pubmed/28881649
http://dx.doi.org/10.18632/oncotarget.17261
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