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A cryptic hydrophobic pocket in the polo-box domain of the polo-like kinase PLK1 regulates substrate recognition and mitotic chromosome segregation

The human polo-like kinase PLK1 coordinates mitotic chromosome segregation by phosphorylating multiple chromatin- and kinetochore-binding proteins. How PLK1 activity is directed to specific substrates via phosphopeptide recognition by its carboxyl-terminal polo-box domain (PBD) is poorly understood....

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Autores principales: Sharma, Pooja, Mahen, Robert, Rossmann, Maxim, Stokes, Jamie E., Hardwick, Bryn, Huggins, David J., Emery, Amy, Kunciw, Dominique L., Hyvönen, Marko, Spring, David R., McKenzie, Grahame J., Venkitaraman, Ashok R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6828814/
https://www.ncbi.nlm.nih.gov/pubmed/31685831
http://dx.doi.org/10.1038/s41598-019-50702-2
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author Sharma, Pooja
Mahen, Robert
Rossmann, Maxim
Stokes, Jamie E.
Hardwick, Bryn
Huggins, David J.
Emery, Amy
Kunciw, Dominique L.
Hyvönen, Marko
Spring, David R.
McKenzie, Grahame J.
Venkitaraman, Ashok R.
author_facet Sharma, Pooja
Mahen, Robert
Rossmann, Maxim
Stokes, Jamie E.
Hardwick, Bryn
Huggins, David J.
Emery, Amy
Kunciw, Dominique L.
Hyvönen, Marko
Spring, David R.
McKenzie, Grahame J.
Venkitaraman, Ashok R.
author_sort Sharma, Pooja
collection PubMed
description The human polo-like kinase PLK1 coordinates mitotic chromosome segregation by phosphorylating multiple chromatin- and kinetochore-binding proteins. How PLK1 activity is directed to specific substrates via phosphopeptide recognition by its carboxyl-terminal polo-box domain (PBD) is poorly understood. Here, we combine molecular, structural and chemical biology to identify a determinant for PLK1 substrate recognition that is essential for proper chromosome segregation. We show that mutations ablating an evolutionarily conserved, Tyr-lined pocket in human PLK1 PBD trigger cellular anomalies in mitotic progression and timing. Tyr pocket mutations selectively impair PLK1 binding to the kinetochore phosphoprotein substrate PBIP1, but not to the centrosomal substrate NEDD1. Through a structure-guided approach, we develop a small-molecule inhibitor, Polotyrin, which occupies the Tyr pocket. Polotyrin recapitulates the mitotic defects caused by mutations in the Tyr pocket, further evidencing its essential function, and exemplifying a new approach for selective PLK1 inhibition. Thus, our findings support a model wherein substrate discrimination via the Tyr pocket in the human PLK1 PBD regulates mitotic chromosome segregation to preserve genome integrity.
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spelling pubmed-68288142019-11-12 A cryptic hydrophobic pocket in the polo-box domain of the polo-like kinase PLK1 regulates substrate recognition and mitotic chromosome segregation Sharma, Pooja Mahen, Robert Rossmann, Maxim Stokes, Jamie E. Hardwick, Bryn Huggins, David J. Emery, Amy Kunciw, Dominique L. Hyvönen, Marko Spring, David R. McKenzie, Grahame J. Venkitaraman, Ashok R. Sci Rep Article The human polo-like kinase PLK1 coordinates mitotic chromosome segregation by phosphorylating multiple chromatin- and kinetochore-binding proteins. How PLK1 activity is directed to specific substrates via phosphopeptide recognition by its carboxyl-terminal polo-box domain (PBD) is poorly understood. Here, we combine molecular, structural and chemical biology to identify a determinant for PLK1 substrate recognition that is essential for proper chromosome segregation. We show that mutations ablating an evolutionarily conserved, Tyr-lined pocket in human PLK1 PBD trigger cellular anomalies in mitotic progression and timing. Tyr pocket mutations selectively impair PLK1 binding to the kinetochore phosphoprotein substrate PBIP1, but not to the centrosomal substrate NEDD1. Through a structure-guided approach, we develop a small-molecule inhibitor, Polotyrin, which occupies the Tyr pocket. Polotyrin recapitulates the mitotic defects caused by mutations in the Tyr pocket, further evidencing its essential function, and exemplifying a new approach for selective PLK1 inhibition. Thus, our findings support a model wherein substrate discrimination via the Tyr pocket in the human PLK1 PBD regulates mitotic chromosome segregation to preserve genome integrity. Nature Publishing Group UK 2019-11-04 /pmc/articles/PMC6828814/ /pubmed/31685831 http://dx.doi.org/10.1038/s41598-019-50702-2 Text en © The Author(s) 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Sharma, Pooja
Mahen, Robert
Rossmann, Maxim
Stokes, Jamie E.
Hardwick, Bryn
Huggins, David J.
Emery, Amy
Kunciw, Dominique L.
Hyvönen, Marko
Spring, David R.
McKenzie, Grahame J.
Venkitaraman, Ashok R.
A cryptic hydrophobic pocket in the polo-box domain of the polo-like kinase PLK1 regulates substrate recognition and mitotic chromosome segregation
title A cryptic hydrophobic pocket in the polo-box domain of the polo-like kinase PLK1 regulates substrate recognition and mitotic chromosome segregation
title_full A cryptic hydrophobic pocket in the polo-box domain of the polo-like kinase PLK1 regulates substrate recognition and mitotic chromosome segregation
title_fullStr A cryptic hydrophobic pocket in the polo-box domain of the polo-like kinase PLK1 regulates substrate recognition and mitotic chromosome segregation
title_full_unstemmed A cryptic hydrophobic pocket in the polo-box domain of the polo-like kinase PLK1 regulates substrate recognition and mitotic chromosome segregation
title_short A cryptic hydrophobic pocket in the polo-box domain of the polo-like kinase PLK1 regulates substrate recognition and mitotic chromosome segregation
title_sort cryptic hydrophobic pocket in the polo-box domain of the polo-like kinase plk1 regulates substrate recognition and mitotic chromosome segregation
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6828814/
https://www.ncbi.nlm.nih.gov/pubmed/31685831
http://dx.doi.org/10.1038/s41598-019-50702-2
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