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Cdk11 is a RanGTP-dependent microtubule stabilization factor that regulates spindle assembly rate

Production of Ran–guanosine triphosphate (GTP) around chromosomes induces local nucleation and plus end stabilization of microtubules (MTs). The nuclear protein TPX2 is required for RanGTP-dependent MT nucleation. To find the MT stabilizer, we affinity purify nuclear localization signal (NLS)–contai...

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Autores principales: Yokoyama, Hideki, Gruss, Oliver J., Rybina, Sofia, Caudron, Maïwen, Schelder, Malgorzata, Wilm, Matthias, Mattaj, Iain W., Karsenti, Eric
Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 2008
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2265385/
https://www.ncbi.nlm.nih.gov/pubmed/18316407
http://dx.doi.org/10.1083/jcb.200706189
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author Yokoyama, Hideki
Gruss, Oliver J.
Rybina, Sofia
Caudron, Maïwen
Schelder, Malgorzata
Wilm, Matthias
Mattaj, Iain W.
Karsenti, Eric
author_facet Yokoyama, Hideki
Gruss, Oliver J.
Rybina, Sofia
Caudron, Maïwen
Schelder, Malgorzata
Wilm, Matthias
Mattaj, Iain W.
Karsenti, Eric
author_sort Yokoyama, Hideki
collection PubMed
description Production of Ran–guanosine triphosphate (GTP) around chromosomes induces local nucleation and plus end stabilization of microtubules (MTs). The nuclear protein TPX2 is required for RanGTP-dependent MT nucleation. To find the MT stabilizer, we affinity purify nuclear localization signal (NLS)–containing proteins from Xenopus laevis egg extracts. This NLS protein fraction contains the MT stabilization activity. After further purification, we used mass spectrometry to identify proteins in active fractions, including cyclin-dependent kinase 11 (Cdk11). Cdk11 localizes on spindle poles and MTs in Xenopus culture cells and egg extracts. Recombinant Cdk11 demonstrates RanGTP-dependent MT stabilization activity, whereas a kinase-dead mutant does not. Inactivation of Cdk11 in egg extracts blocks RanGTP-dependent MT stabilization and dramatically decreases the spindle assembly rate. Simultaneous depletion of TPX2 completely inhibits centrosome-dependent spindle assembly. Our results indicate that Cdk11 is responsible for RanGTP-dependent MT stabilization around chromosomes and that this local stabilization is essential for normal rates of spindle assembly and spindle function.
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spelling pubmed-22653852008-09-10 Cdk11 is a RanGTP-dependent microtubule stabilization factor that regulates spindle assembly rate Yokoyama, Hideki Gruss, Oliver J. Rybina, Sofia Caudron, Maïwen Schelder, Malgorzata Wilm, Matthias Mattaj, Iain W. Karsenti, Eric J Cell Biol Research Articles Production of Ran–guanosine triphosphate (GTP) around chromosomes induces local nucleation and plus end stabilization of microtubules (MTs). The nuclear protein TPX2 is required for RanGTP-dependent MT nucleation. To find the MT stabilizer, we affinity purify nuclear localization signal (NLS)–containing proteins from Xenopus laevis egg extracts. This NLS protein fraction contains the MT stabilization activity. After further purification, we used mass spectrometry to identify proteins in active fractions, including cyclin-dependent kinase 11 (Cdk11). Cdk11 localizes on spindle poles and MTs in Xenopus culture cells and egg extracts. Recombinant Cdk11 demonstrates RanGTP-dependent MT stabilization activity, whereas a kinase-dead mutant does not. Inactivation of Cdk11 in egg extracts blocks RanGTP-dependent MT stabilization and dramatically decreases the spindle assembly rate. Simultaneous depletion of TPX2 completely inhibits centrosome-dependent spindle assembly. Our results indicate that Cdk11 is responsible for RanGTP-dependent MT stabilization around chromosomes and that this local stabilization is essential for normal rates of spindle assembly and spindle function. The Rockefeller University Press 2008-03-10 /pmc/articles/PMC2265385/ /pubmed/18316407 http://dx.doi.org/10.1083/jcb.200706189 Text en Copyright © 2008, The Rockefeller University Press This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).
spellingShingle Research Articles
Yokoyama, Hideki
Gruss, Oliver J.
Rybina, Sofia
Caudron, Maïwen
Schelder, Malgorzata
Wilm, Matthias
Mattaj, Iain W.
Karsenti, Eric
Cdk11 is a RanGTP-dependent microtubule stabilization factor that regulates spindle assembly rate
title Cdk11 is a RanGTP-dependent microtubule stabilization factor that regulates spindle assembly rate
title_full Cdk11 is a RanGTP-dependent microtubule stabilization factor that regulates spindle assembly rate
title_fullStr Cdk11 is a RanGTP-dependent microtubule stabilization factor that regulates spindle assembly rate
title_full_unstemmed Cdk11 is a RanGTP-dependent microtubule stabilization factor that regulates spindle assembly rate
title_short Cdk11 is a RanGTP-dependent microtubule stabilization factor that regulates spindle assembly rate
title_sort cdk11 is a rangtp-dependent microtubule stabilization factor that regulates spindle assembly rate
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2265385/
https://www.ncbi.nlm.nih.gov/pubmed/18316407
http://dx.doi.org/10.1083/jcb.200706189
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