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Multisite phosphorylation of C-Nap1 releases it from Cep135 to trigger centrosome disjunction
During mitotic entry, centrosomes separate to establish the bipolar spindle. Delays in centrosome separation can perturb chromosome segregation and promote genetic instability. However, interphase centrosomes are physically tethered by a proteinaceous linker composed of C-Nap1 (also known as CEP250)...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Company of Biologists
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4038944/ https://www.ncbi.nlm.nih.gov/pubmed/24695856 http://dx.doi.org/10.1242/jcs.142331 |
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author | Hardy, Tara Lee, Miseon Hames, Rebecca S. Prosser, Suzanna L. Cheary, Donna-Marie Samant, Mugdha D. Schultz, Francisca Baxter, Joanne E. Rhee, Kunsoo Fry, Andrew M. |
author_facet | Hardy, Tara Lee, Miseon Hames, Rebecca S. Prosser, Suzanna L. Cheary, Donna-Marie Samant, Mugdha D. Schultz, Francisca Baxter, Joanne E. Rhee, Kunsoo Fry, Andrew M. |
author_sort | Hardy, Tara |
collection | PubMed |
description | During mitotic entry, centrosomes separate to establish the bipolar spindle. Delays in centrosome separation can perturb chromosome segregation and promote genetic instability. However, interphase centrosomes are physically tethered by a proteinaceous linker composed of C-Nap1 (also known as CEP250) and the filamentous protein rootletin. Linker disassembly occurs at the onset of mitosis in a process known as centrosome disjunction and is triggered by the Nek2-dependent phosphorylation of C-Nap1. However, the mechanistic consequences of C-Nap1 phosphorylation are unknown. Here, we demonstrate that Nek2 phosphorylates multiple residues within the C-terminal domain of C-Nap1 and, collectively, these phosphorylation events lead to loss of oligomerization and centrosome association. Mutations in non-phosphorylatable residues that make the domain more acidic are sufficient to release C-Nap1 from the centrosome, suggesting that it is an increase in overall negative charge that is required for this process. Importantly, phosphorylation of C-Nap1 also perturbs interaction with the core centriolar protein, Cep135, and interaction of endogenous C-Nap1 and Cep135 proteins is specifically lost in mitosis. We therefore propose that multisite phosphorylation of C-Nap1 by Nek2 perturbs both oligomerization and Cep135 interaction, and this precipitates centrosome disjunction at the onset of mitosis. |
format | Online Article Text |
id | pubmed-4038944 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | The Company of Biologists |
record_format | MEDLINE/PubMed |
spelling | pubmed-40389442014-06-12 Multisite phosphorylation of C-Nap1 releases it from Cep135 to trigger centrosome disjunction Hardy, Tara Lee, Miseon Hames, Rebecca S. Prosser, Suzanna L. Cheary, Donna-Marie Samant, Mugdha D. Schultz, Francisca Baxter, Joanne E. Rhee, Kunsoo Fry, Andrew M. J Cell Sci Research Article During mitotic entry, centrosomes separate to establish the bipolar spindle. Delays in centrosome separation can perturb chromosome segregation and promote genetic instability. However, interphase centrosomes are physically tethered by a proteinaceous linker composed of C-Nap1 (also known as CEP250) and the filamentous protein rootletin. Linker disassembly occurs at the onset of mitosis in a process known as centrosome disjunction and is triggered by the Nek2-dependent phosphorylation of C-Nap1. However, the mechanistic consequences of C-Nap1 phosphorylation are unknown. Here, we demonstrate that Nek2 phosphorylates multiple residues within the C-terminal domain of C-Nap1 and, collectively, these phosphorylation events lead to loss of oligomerization and centrosome association. Mutations in non-phosphorylatable residues that make the domain more acidic are sufficient to release C-Nap1 from the centrosome, suggesting that it is an increase in overall negative charge that is required for this process. Importantly, phosphorylation of C-Nap1 also perturbs interaction with the core centriolar protein, Cep135, and interaction of endogenous C-Nap1 and Cep135 proteins is specifically lost in mitosis. We therefore propose that multisite phosphorylation of C-Nap1 by Nek2 perturbs both oligomerization and Cep135 interaction, and this precipitates centrosome disjunction at the onset of mitosis. The Company of Biologists 2014-06-01 /pmc/articles/PMC4038944/ /pubmed/24695856 http://dx.doi.org/10.1242/jcs.142331 Text en © 2014. 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 Hardy, Tara Lee, Miseon Hames, Rebecca S. Prosser, Suzanna L. Cheary, Donna-Marie Samant, Mugdha D. Schultz, Francisca Baxter, Joanne E. Rhee, Kunsoo Fry, Andrew M. Multisite phosphorylation of C-Nap1 releases it from Cep135 to trigger centrosome disjunction |
title | Multisite phosphorylation of C-Nap1 releases it from Cep135 to trigger centrosome disjunction |
title_full | Multisite phosphorylation of C-Nap1 releases it from Cep135 to trigger centrosome disjunction |
title_fullStr | Multisite phosphorylation of C-Nap1 releases it from Cep135 to trigger centrosome disjunction |
title_full_unstemmed | Multisite phosphorylation of C-Nap1 releases it from Cep135 to trigger centrosome disjunction |
title_short | Multisite phosphorylation of C-Nap1 releases it from Cep135 to trigger centrosome disjunction |
title_sort | multisite phosphorylation of c-nap1 releases it from cep135 to trigger centrosome disjunction |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4038944/ https://www.ncbi.nlm.nih.gov/pubmed/24695856 http://dx.doi.org/10.1242/jcs.142331 |
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