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Requirement for PLK1 kinase activity in the maintenance of a robust spindle assembly checkpoint

During mitotic arrest induced by microtubule targeting drugs, the weakening of the spindle assembly checkpoint (SAC) allows cells to progress through the cell cycle without chromosome segregation occurring. PLK1 kinase plays a major role in mitosis and emerging evidence indicates that PLK1 is also i...

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Autores principales: O'Connor, Aisling, Maffini, Stefano, Rainey, Michael D., Kaczmarczyk, Agnieszka, Gaboriau, David, Musacchio, Andrea, Santocanale, Corrado
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Company of Biologists Ltd 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4728306/
https://www.ncbi.nlm.nih.gov/pubmed/26685311
http://dx.doi.org/10.1242/bio.014969
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author O'Connor, Aisling
Maffini, Stefano
Rainey, Michael D.
Kaczmarczyk, Agnieszka
Gaboriau, David
Musacchio, Andrea
Santocanale, Corrado
author_facet O'Connor, Aisling
Maffini, Stefano
Rainey, Michael D.
Kaczmarczyk, Agnieszka
Gaboriau, David
Musacchio, Andrea
Santocanale, Corrado
author_sort O'Connor, Aisling
collection PubMed
description During mitotic arrest induced by microtubule targeting drugs, the weakening of the spindle assembly checkpoint (SAC) allows cells to progress through the cell cycle without chromosome segregation occurring. PLK1 kinase plays a major role in mitosis and emerging evidence indicates that PLK1 is also involved in establishing the checkpoint and maintaining SAC signalling. However, mechanistically, the role of PLK1 in the SAC is not fully understood, with several recent reports indicating that it can cooperate with either one of the major checkpoint kinases, Aurora B or MPS1. In this study, we assess the role of PLK1 in SAC maintenance. We find that in nocodazole-arrested U2OS cells, PLK1 activity is continuously required for maintaining Aurora B protein localisation and activity at kinetochores. Consistent with published data we find that upon PLK1 inhibition, phosphoThr3-H3, a marker of Haspin activity, is reduced. Intriguingly, Aurora B inhibition causes PLK1 to relocalise from kinetochores into fewer and much larger foci, possibly due to incomplete recruitment of outer kinetochore proteins. Importantly, PLK1 inhibition, together with partial inhibition of Aurora B, allows efficient SAC override to occur. This phenotype is more pronounced than the phenotype observed by combining the same PLK1 inhibitors with partial MPS1 inhibition. We also find that PLK1 inhibition does not obviously cooperate with Haspin inhibition to promote SAC override. These results indicate that PLK1 is directly involved in maintaining efficient SAC signalling, possibly by cooperating in a positive feedback loop with Aurora B, and that partially redundant mechanisms exist which reinforce the SAC.
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spelling pubmed-47283062016-02-01 Requirement for PLK1 kinase activity in the maintenance of a robust spindle assembly checkpoint O'Connor, Aisling Maffini, Stefano Rainey, Michael D. Kaczmarczyk, Agnieszka Gaboriau, David Musacchio, Andrea Santocanale, Corrado Biol Open Research Article During mitotic arrest induced by microtubule targeting drugs, the weakening of the spindle assembly checkpoint (SAC) allows cells to progress through the cell cycle without chromosome segregation occurring. PLK1 kinase plays a major role in mitosis and emerging evidence indicates that PLK1 is also involved in establishing the checkpoint and maintaining SAC signalling. However, mechanistically, the role of PLK1 in the SAC is not fully understood, with several recent reports indicating that it can cooperate with either one of the major checkpoint kinases, Aurora B or MPS1. In this study, we assess the role of PLK1 in SAC maintenance. We find that in nocodazole-arrested U2OS cells, PLK1 activity is continuously required for maintaining Aurora B protein localisation and activity at kinetochores. Consistent with published data we find that upon PLK1 inhibition, phosphoThr3-H3, a marker of Haspin activity, is reduced. Intriguingly, Aurora B inhibition causes PLK1 to relocalise from kinetochores into fewer and much larger foci, possibly due to incomplete recruitment of outer kinetochore proteins. Importantly, PLK1 inhibition, together with partial inhibition of Aurora B, allows efficient SAC override to occur. This phenotype is more pronounced than the phenotype observed by combining the same PLK1 inhibitors with partial MPS1 inhibition. We also find that PLK1 inhibition does not obviously cooperate with Haspin inhibition to promote SAC override. These results indicate that PLK1 is directly involved in maintaining efficient SAC signalling, possibly by cooperating in a positive feedback loop with Aurora B, and that partially redundant mechanisms exist which reinforce the SAC. The Company of Biologists Ltd 2015-12-18 /pmc/articles/PMC4728306/ /pubmed/26685311 http://dx.doi.org/10.1242/bio.014969 Text en © 2016. Published by The Company of Biologists Ltd 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), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.
spellingShingle Research Article
O'Connor, Aisling
Maffini, Stefano
Rainey, Michael D.
Kaczmarczyk, Agnieszka
Gaboriau, David
Musacchio, Andrea
Santocanale, Corrado
Requirement for PLK1 kinase activity in the maintenance of a robust spindle assembly checkpoint
title Requirement for PLK1 kinase activity in the maintenance of a robust spindle assembly checkpoint
title_full Requirement for PLK1 kinase activity in the maintenance of a robust spindle assembly checkpoint
title_fullStr Requirement for PLK1 kinase activity in the maintenance of a robust spindle assembly checkpoint
title_full_unstemmed Requirement for PLK1 kinase activity in the maintenance of a robust spindle assembly checkpoint
title_short Requirement for PLK1 kinase activity in the maintenance of a robust spindle assembly checkpoint
title_sort requirement for plk1 kinase activity in the maintenance of a robust spindle assembly checkpoint
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4728306/
https://www.ncbi.nlm.nih.gov/pubmed/26685311
http://dx.doi.org/10.1242/bio.014969
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