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Polo-like kinase phosphorylation determines Caenorhabditis elegans centrosome size and density by biasing SPD-5 toward an assembly-competent conformation
Centrosomes are major microtubule-organizing centers composed of centrioles surrounded by an extensive proteinacious layer called the pericentriolar material (PCM). In Caenorhabditis elegans embryos, the mitotic PCM expands by Polo-like kinase 1 (PLK-1) phosphorylation-accelerated assembly of SPD-5...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Company of Biologists Ltd
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5087692/ https://www.ncbi.nlm.nih.gov/pubmed/27591191 http://dx.doi.org/10.1242/bio.020990 |
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author | Wueseke, Oliver Zwicker, David Schwager, Anne Wong, Yao Liang Oegema, Karen Jülicher, Frank Hyman, Anthony A. Woodruff, Jeffrey B. |
author_facet | Wueseke, Oliver Zwicker, David Schwager, Anne Wong, Yao Liang Oegema, Karen Jülicher, Frank Hyman, Anthony A. Woodruff, Jeffrey B. |
author_sort | Wueseke, Oliver |
collection | PubMed |
description | Centrosomes are major microtubule-organizing centers composed of centrioles surrounded by an extensive proteinacious layer called the pericentriolar material (PCM). In Caenorhabditis elegans embryos, the mitotic PCM expands by Polo-like kinase 1 (PLK-1) phosphorylation-accelerated assembly of SPD-5 molecules into supramolecular scaffolds. However, how PLK-1 phosphorylation regulates SPD-5 assembly is not known. We found that a mutant version of SPD-5 that is insensitive to PLK-1 phosphorylation (SPD-5(4A)) could localize to PCM but was unable to rescue the reduction in PCM size and density when wild-type SPD-5 levels were decreased. In vitro, purified SPD-5(4A) self-assembled into functional supramolecular scaffolds over long time scales, suggesting that phosphorylation only controls the rate of SPD-5 scaffold assembly. Furthermore, the SPD-5 scaffold, once assembled, remained intact and supported microtubule nucleation in the absence of PLK-1 activity in vivo. We conclude that PLK-1 is required for rapid assembly of the PCM scaffold but not for scaffold maintenance or function. Based on this idea, we developed a theoretical model that adequately predicted PCM growth rates in different mutant conditions in vivo. We propose that PLK-1 phosphorylation-dependent conversion of SPD-5 into an assembly-competent form underlies PCM formation in vivo and that the rate of this conversion determines final PCM size and density. |
format | Online Article Text |
id | pubmed-5087692 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | The Company of Biologists Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-50876922016-10-31 Polo-like kinase phosphorylation determines Caenorhabditis elegans centrosome size and density by biasing SPD-5 toward an assembly-competent conformation Wueseke, Oliver Zwicker, David Schwager, Anne Wong, Yao Liang Oegema, Karen Jülicher, Frank Hyman, Anthony A. Woodruff, Jeffrey B. Biol Open Research Article Centrosomes are major microtubule-organizing centers composed of centrioles surrounded by an extensive proteinacious layer called the pericentriolar material (PCM). In Caenorhabditis elegans embryos, the mitotic PCM expands by Polo-like kinase 1 (PLK-1) phosphorylation-accelerated assembly of SPD-5 molecules into supramolecular scaffolds. However, how PLK-1 phosphorylation regulates SPD-5 assembly is not known. We found that a mutant version of SPD-5 that is insensitive to PLK-1 phosphorylation (SPD-5(4A)) could localize to PCM but was unable to rescue the reduction in PCM size and density when wild-type SPD-5 levels were decreased. In vitro, purified SPD-5(4A) self-assembled into functional supramolecular scaffolds over long time scales, suggesting that phosphorylation only controls the rate of SPD-5 scaffold assembly. Furthermore, the SPD-5 scaffold, once assembled, remained intact and supported microtubule nucleation in the absence of PLK-1 activity in vivo. We conclude that PLK-1 is required for rapid assembly of the PCM scaffold but not for scaffold maintenance or function. Based on this idea, we developed a theoretical model that adequately predicted PCM growth rates in different mutant conditions in vivo. We propose that PLK-1 phosphorylation-dependent conversion of SPD-5 into an assembly-competent form underlies PCM formation in vivo and that the rate of this conversion determines final PCM size and density. The Company of Biologists Ltd 2016-09-02 /pmc/articles/PMC5087692/ /pubmed/27591191 http://dx.doi.org/10.1242/bio.020990 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 Wueseke, Oliver Zwicker, David Schwager, Anne Wong, Yao Liang Oegema, Karen Jülicher, Frank Hyman, Anthony A. Woodruff, Jeffrey B. Polo-like kinase phosphorylation determines Caenorhabditis elegans centrosome size and density by biasing SPD-5 toward an assembly-competent conformation |
title | Polo-like kinase phosphorylation determines Caenorhabditis
elegans centrosome size and density by biasing SPD-5 toward an assembly-competent conformation |
title_full | Polo-like kinase phosphorylation determines Caenorhabditis
elegans centrosome size and density by biasing SPD-5 toward an assembly-competent conformation |
title_fullStr | Polo-like kinase phosphorylation determines Caenorhabditis
elegans centrosome size and density by biasing SPD-5 toward an assembly-competent conformation |
title_full_unstemmed | Polo-like kinase phosphorylation determines Caenorhabditis
elegans centrosome size and density by biasing SPD-5 toward an assembly-competent conformation |
title_short | Polo-like kinase phosphorylation determines Caenorhabditis
elegans centrosome size and density by biasing SPD-5 toward an assembly-competent conformation |
title_sort | polo-like kinase phosphorylation determines caenorhabditis
elegans centrosome size and density by biasing spd-5 toward an assembly-competent conformation |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5087692/ https://www.ncbi.nlm.nih.gov/pubmed/27591191 http://dx.doi.org/10.1242/bio.020990 |
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