A cryptic hydrophobic pocket in the polo-box domain of the polo-like kinase PLK1 regulates substrate recognition and mitotic chromosome segregation
| Autor: | Pooja Sharma, Marko Hyvönen, Robert Mahen, Bryn Hardwick, David J. Huggins, David R. Spring, Ashok R. Venkitaraman, M. Rossmann, Jamie E. Stokes, Amy Emery, D.L. Kunciw, Grahame J. McKenzie |
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| Přispěvatelé: | Apollo - University of Cambridge Repository, Rossmann, Maxim [0000-0001-8811-3277], Huggins, David [0000-0003-1579-2496], Spring, David [0000-0001-7355-2824] |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
82/29 NEDD1 lcsh:Medicine Cell Cycle Proteins Polo-like kinase 01 natural sciences Substrate Specificity Chromosome segregation Histones Chromosome Segregation RNA Small Interfering 14/19 lcsh:Science Kinetochores 3' Untranslated Regions 13/89 45/70 Multidisciplinary Kinetochore Chemistry Small molecules article 45/77 Cell biology Chromatin 3. Good health 631/92/275 RNA Interference 631/92/612/1246 Protein Binding Cell Survival 631/80/103/1966 145 Mitosis Kinases 13/106 13/109 Protein Serine-Threonine Kinases PLK1 631/92/613 03 medical and health sciences Protein Domains Proto-Oncogene Proteins Humans Protein Kinase Inhibitors 14/35 010405 organic chemistry 631/80/641/1655 lcsh:R Phosphoproteins 0104 chemical sciences Protein Structure Tertiary 030104 developmental biology Mutagenesis Phosphoprotein 14/63 lcsh:Q 82/51 HeLa Cells |
| Zdroj: | Scientific Reports Scientific Reports, Vol 9, Iss 1, Pp 1-15 (2019) |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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