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ATP-competitive Plk1 inhibitors induce caspase 3-mediated Plk1 cleavage and activation in hematopoietic cell lines

Polo-like kinases (Plks) define a highly conserved family of Ser/Thr kinases with crucial roles in the regulation of cell division. Here we show that Plk1 is cleaved by caspase 3, but not by other caspases in different hematopoietic cell lines treated with competitive inhibitors of the ATP-binding p...

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Autores principales: Dufies, Maeva, Ambrosetti, Damien, Boulakirba, Sonia, Calleja, Anne, Savy, Coline, Furstoss, Nathan, Zerhouni, Marwa, Parola, Julien, Aira-Diaz, Lazaro, Marchetti, Sandrine, Orange, Francois, Lacas-Gervais, Sandra, Luciano, Frederic, Jacquel, Arnaud, Robert, Guillaume, Pagès, Gilles, Auberger, Patrick
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Impact Journals LLC 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5834281/
https://www.ncbi.nlm.nih.gov/pubmed/29541386
http://dx.doi.org/10.18632/oncotarget.23650
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author Dufies, Maeva
Ambrosetti, Damien
Boulakirba, Sonia
Calleja, Anne
Savy, Coline
Furstoss, Nathan
Zerhouni, Marwa
Parola, Julien
Aira-Diaz, Lazaro
Marchetti, Sandrine
Orange, Francois
Lacas-Gervais, Sandra
Luciano, Frederic
Jacquel, Arnaud
Robert, Guillaume
Pagès, Gilles
Auberger, Patrick
author_facet Dufies, Maeva
Ambrosetti, Damien
Boulakirba, Sonia
Calleja, Anne
Savy, Coline
Furstoss, Nathan
Zerhouni, Marwa
Parola, Julien
Aira-Diaz, Lazaro
Marchetti, Sandrine
Orange, Francois
Lacas-Gervais, Sandra
Luciano, Frederic
Jacquel, Arnaud
Robert, Guillaume
Pagès, Gilles
Auberger, Patrick
author_sort Dufies, Maeva
collection PubMed
description Polo-like kinases (Plks) define a highly conserved family of Ser/Thr kinases with crucial roles in the regulation of cell division. Here we show that Plk1 is cleaved by caspase 3, but not by other caspases in different hematopoietic cell lines treated with competitive inhibitors of the ATP-binding pocket of Plk1. Intriguingly, Plk1 was not cleaved in cells treated with Rigosertib, a non-competitive inhibitor of Plk1, suggesting that binding of the inhibitor to the ATP binding pocket of Plk1 triggers a conformational change and unmasks a cryptic caspase 3 cleavage site on the protein. Cleavage occurs after Asp-404 in a DYSD/K sequence and separates the kinase domain from the two PBDs of Plk1. All Plk1 inhibitors triggered G2/M arrest, activation of caspases 2 and 3, polyploidy, multiple nuclei and mitotic catastrophe, albeit at higher concentrations in the case of Rigosertib. Upon BI-2536 treatment, Plk1 cleavage occurred only in the cytosolic fraction and cleaved Plk1 accumulated in this subcellular compartment. Importantly, the cleaved N-Terminal fragment of Plk1 exhibited a higher enzymatic activity than its non-cleaved counterpart and accumulated into the cytoplasm conversely to the full length and the C-Terminal Plk1 fragments that were found essentially into the nucleus. Finally, the DYSD/K cleavage site was highly conserved during evolution from c. elegans to human. In conclusion, we described herein for the first time a specific cleavage of Plk1 by caspase 3 following treatment of cancer cells with ATP-competitive inhibitors of Plk1.
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spelling pubmed-58342812018-03-14 ATP-competitive Plk1 inhibitors induce caspase 3-mediated Plk1 cleavage and activation in hematopoietic cell lines Dufies, Maeva Ambrosetti, Damien Boulakirba, Sonia Calleja, Anne Savy, Coline Furstoss, Nathan Zerhouni, Marwa Parola, Julien Aira-Diaz, Lazaro Marchetti, Sandrine Orange, Francois Lacas-Gervais, Sandra Luciano, Frederic Jacquel, Arnaud Robert, Guillaume Pagès, Gilles Auberger, Patrick Oncotarget Research Paper Polo-like kinases (Plks) define a highly conserved family of Ser/Thr kinases with crucial roles in the regulation of cell division. Here we show that Plk1 is cleaved by caspase 3, but not by other caspases in different hematopoietic cell lines treated with competitive inhibitors of the ATP-binding pocket of Plk1. Intriguingly, Plk1 was not cleaved in cells treated with Rigosertib, a non-competitive inhibitor of Plk1, suggesting that binding of the inhibitor to the ATP binding pocket of Plk1 triggers a conformational change and unmasks a cryptic caspase 3 cleavage site on the protein. Cleavage occurs after Asp-404 in a DYSD/K sequence and separates the kinase domain from the two PBDs of Plk1. All Plk1 inhibitors triggered G2/M arrest, activation of caspases 2 and 3, polyploidy, multiple nuclei and mitotic catastrophe, albeit at higher concentrations in the case of Rigosertib. Upon BI-2536 treatment, Plk1 cleavage occurred only in the cytosolic fraction and cleaved Plk1 accumulated in this subcellular compartment. Importantly, the cleaved N-Terminal fragment of Plk1 exhibited a higher enzymatic activity than its non-cleaved counterpart and accumulated into the cytoplasm conversely to the full length and the C-Terminal Plk1 fragments that were found essentially into the nucleus. Finally, the DYSD/K cleavage site was highly conserved during evolution from c. elegans to human. In conclusion, we described herein for the first time a specific cleavage of Plk1 by caspase 3 following treatment of cancer cells with ATP-competitive inhibitors of Plk1. Impact Journals LLC 2017-12-23 /pmc/articles/PMC5834281/ /pubmed/29541386 http://dx.doi.org/10.18632/oncotarget.23650 Text en Copyright: © 2018 Dufies 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 (http://creativecommons.org/licenses/by/3.0/) 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
Dufies, Maeva
Ambrosetti, Damien
Boulakirba, Sonia
Calleja, Anne
Savy, Coline
Furstoss, Nathan
Zerhouni, Marwa
Parola, Julien
Aira-Diaz, Lazaro
Marchetti, Sandrine
Orange, Francois
Lacas-Gervais, Sandra
Luciano, Frederic
Jacquel, Arnaud
Robert, Guillaume
Pagès, Gilles
Auberger, Patrick
ATP-competitive Plk1 inhibitors induce caspase 3-mediated Plk1 cleavage and activation in hematopoietic cell lines
title ATP-competitive Plk1 inhibitors induce caspase 3-mediated Plk1 cleavage and activation in hematopoietic cell lines
title_full ATP-competitive Plk1 inhibitors induce caspase 3-mediated Plk1 cleavage and activation in hematopoietic cell lines
title_fullStr ATP-competitive Plk1 inhibitors induce caspase 3-mediated Plk1 cleavage and activation in hematopoietic cell lines
title_full_unstemmed ATP-competitive Plk1 inhibitors induce caspase 3-mediated Plk1 cleavage and activation in hematopoietic cell lines
title_short ATP-competitive Plk1 inhibitors induce caspase 3-mediated Plk1 cleavage and activation in hematopoietic cell lines
title_sort atp-competitive plk1 inhibitors induce caspase 3-mediated plk1 cleavage and activation in hematopoietic cell lines
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5834281/
https://www.ncbi.nlm.nih.gov/pubmed/29541386
http://dx.doi.org/10.18632/oncotarget.23650
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