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Systematic kinetic analysis of mitotic dis- and reassembly of the nuclear pore in living cells

During mitosis in higher eukaryotes, nuclear pore complexes (NPCs) disassemble in prophase and are rebuilt in anaphase and telophase. NPC formation is hypothesized to occur by the interaction of mitotically stable subcomplexes that form defined structural intermediates. To determine the sequence of...

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Autores principales: Dultz, Elisa, Zanin, Esther, Wurzenberger, Claudia, Braun, Marion, Rabut, Gwénaël, Sironi, Lucia, Ellenberg, Jan
Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 2008
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2265396/
https://www.ncbi.nlm.nih.gov/pubmed/18316408
http://dx.doi.org/10.1083/jcb.200707026
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author Dultz, Elisa
Zanin, Esther
Wurzenberger, Claudia
Braun, Marion
Rabut, Gwénaël
Sironi, Lucia
Ellenberg, Jan
author_facet Dultz, Elisa
Zanin, Esther
Wurzenberger, Claudia
Braun, Marion
Rabut, Gwénaël
Sironi, Lucia
Ellenberg, Jan
author_sort Dultz, Elisa
collection PubMed
description During mitosis in higher eukaryotes, nuclear pore complexes (NPCs) disassemble in prophase and are rebuilt in anaphase and telophase. NPC formation is hypothesized to occur by the interaction of mitotically stable subcomplexes that form defined structural intermediates. To determine the sequence of events that lead to breakdown and reformation of functional NPCs during mitosis, we present here our quantitative assay based on confocal time-lapse microscopy of single dividing cells. We use this assay to systematically investigate the kinetics of dis- and reassembly for eight nucleoporin subcomplexes relative to nuclear transport in NRK cells, linking the assembly state of the NPC with its function. Our data establish that NPC assembly is an ordered stepwise process that leads to import function already in a partially assembled state. We furthermore find that nucleoporin dissociation does not occur in the reverse order from binding during assembly, which may indicate a distinct mechanism.
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spelling pubmed-22653962008-09-10 Systematic kinetic analysis of mitotic dis- and reassembly of the nuclear pore in living cells Dultz, Elisa Zanin, Esther Wurzenberger, Claudia Braun, Marion Rabut, Gwénaël Sironi, Lucia Ellenberg, Jan J Cell Biol Research Articles During mitosis in higher eukaryotes, nuclear pore complexes (NPCs) disassemble in prophase and are rebuilt in anaphase and telophase. NPC formation is hypothesized to occur by the interaction of mitotically stable subcomplexes that form defined structural intermediates. To determine the sequence of events that lead to breakdown and reformation of functional NPCs during mitosis, we present here our quantitative assay based on confocal time-lapse microscopy of single dividing cells. We use this assay to systematically investigate the kinetics of dis- and reassembly for eight nucleoporin subcomplexes relative to nuclear transport in NRK cells, linking the assembly state of the NPC with its function. Our data establish that NPC assembly is an ordered stepwise process that leads to import function already in a partially assembled state. We furthermore find that nucleoporin dissociation does not occur in the reverse order from binding during assembly, which may indicate a distinct mechanism. The Rockefeller University Press 2008-03-10 /pmc/articles/PMC2265396/ /pubmed/18316408 http://dx.doi.org/10.1083/jcb.200707026 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
Dultz, Elisa
Zanin, Esther
Wurzenberger, Claudia
Braun, Marion
Rabut, Gwénaël
Sironi, Lucia
Ellenberg, Jan
Systematic kinetic analysis of mitotic dis- and reassembly of the nuclear pore in living cells
title Systematic kinetic analysis of mitotic dis- and reassembly of the nuclear pore in living cells
title_full Systematic kinetic analysis of mitotic dis- and reassembly of the nuclear pore in living cells
title_fullStr Systematic kinetic analysis of mitotic dis- and reassembly of the nuclear pore in living cells
title_full_unstemmed Systematic kinetic analysis of mitotic dis- and reassembly of the nuclear pore in living cells
title_short Systematic kinetic analysis of mitotic dis- and reassembly of the nuclear pore in living cells
title_sort systematic kinetic analysis of mitotic dis- and reassembly of the nuclear pore in living cells
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2265396/
https://www.ncbi.nlm.nih.gov/pubmed/18316408
http://dx.doi.org/10.1083/jcb.200707026
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