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Ran-dependent docking of importin-β to RanBP2/Nup358 filaments is essential for protein import and cell viability
RanBP2/Nup358, the major component of the cytoplasmic filaments of the nuclear pore complex (NPC), is essential for mouse embryogenesis and is implicated in both macromolecular transport and mitosis, but its specific molecular functions are unknown. Using RanBP2 conditional knockout mouse embryonic...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Rockefeller University Press
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3160583/ https://www.ncbi.nlm.nih.gov/pubmed/21859863 http://dx.doi.org/10.1083/jcb.201102018 |
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author | Hamada, Masakazu Haeger, Anna Jeganathan, Karthik B. van Ree, Janine H. Malureanu, Liviu Wälde, Sarah Joseph, Jomon Kehlenbach, Ralph H. van Deursen, Jan M. |
author_facet | Hamada, Masakazu Haeger, Anna Jeganathan, Karthik B. van Ree, Janine H. Malureanu, Liviu Wälde, Sarah Joseph, Jomon Kehlenbach, Ralph H. van Deursen, Jan M. |
author_sort | Hamada, Masakazu |
collection | PubMed |
description | RanBP2/Nup358, the major component of the cytoplasmic filaments of the nuclear pore complex (NPC), is essential for mouse embryogenesis and is implicated in both macromolecular transport and mitosis, but its specific molecular functions are unknown. Using RanBP2 conditional knockout mouse embryonic fibroblasts and a series of mutant constructs, we show that transport, rather than mitotic, functions of RanBP2 are required for cell viability. Cre-mediated RanBP2 inactivation caused cell death with defects in M9- and classical nuclear localization signal (cNLS)–mediated protein import, nuclear export signal–mediated protein export, and messenger ribonucleic acid export but no apparent mitotic failure. A short N-terminal RanBP2 fragment harboring the NPC-binding domain, three phenylalanine-glycine motifs, and one Ran-binding domain (RBD) corrected all transport defects and restored viability. Mutation of the RBD within this fragment caused lethality and perturbed binding to Ran guanosine triphosphate (GTP)–importin-β, accumulation of importin-β at nuclear pores, and cNLS-mediated protein import. These data suggest that a critical function of RanBP2 is to capture recycling RanGTP–importin-β complexes at cytoplasmic fibrils to allow for adequate cNLS-mediated cargo import. |
format | Online Article Text |
id | pubmed-3160583 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | The Rockefeller University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-31605832012-02-22 Ran-dependent docking of importin-β to RanBP2/Nup358 filaments is essential for protein import and cell viability Hamada, Masakazu Haeger, Anna Jeganathan, Karthik B. van Ree, Janine H. Malureanu, Liviu Wälde, Sarah Joseph, Jomon Kehlenbach, Ralph H. van Deursen, Jan M. J Cell Biol Research Articles RanBP2/Nup358, the major component of the cytoplasmic filaments of the nuclear pore complex (NPC), is essential for mouse embryogenesis and is implicated in both macromolecular transport and mitosis, but its specific molecular functions are unknown. Using RanBP2 conditional knockout mouse embryonic fibroblasts and a series of mutant constructs, we show that transport, rather than mitotic, functions of RanBP2 are required for cell viability. Cre-mediated RanBP2 inactivation caused cell death with defects in M9- and classical nuclear localization signal (cNLS)–mediated protein import, nuclear export signal–mediated protein export, and messenger ribonucleic acid export but no apparent mitotic failure. A short N-terminal RanBP2 fragment harboring the NPC-binding domain, three phenylalanine-glycine motifs, and one Ran-binding domain (RBD) corrected all transport defects and restored viability. Mutation of the RBD within this fragment caused lethality and perturbed binding to Ran guanosine triphosphate (GTP)–importin-β, accumulation of importin-β at nuclear pores, and cNLS-mediated protein import. These data suggest that a critical function of RanBP2 is to capture recycling RanGTP–importin-β complexes at cytoplasmic fibrils to allow for adequate cNLS-mediated cargo import. The Rockefeller University Press 2011-08-22 /pmc/articles/PMC3160583/ /pubmed/21859863 http://dx.doi.org/10.1083/jcb.201102018 Text en © 2011 Hamada et al. 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 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/). |
spellingShingle | Research Articles Hamada, Masakazu Haeger, Anna Jeganathan, Karthik B. van Ree, Janine H. Malureanu, Liviu Wälde, Sarah Joseph, Jomon Kehlenbach, Ralph H. van Deursen, Jan M. Ran-dependent docking of importin-β to RanBP2/Nup358 filaments is essential for protein import and cell viability |
title | Ran-dependent docking of importin-β to RanBP2/Nup358 filaments is essential for protein import and cell viability |
title_full | Ran-dependent docking of importin-β to RanBP2/Nup358 filaments is essential for protein import and cell viability |
title_fullStr | Ran-dependent docking of importin-β to RanBP2/Nup358 filaments is essential for protein import and cell viability |
title_full_unstemmed | Ran-dependent docking of importin-β to RanBP2/Nup358 filaments is essential for protein import and cell viability |
title_short | Ran-dependent docking of importin-β to RanBP2/Nup358 filaments is essential for protein import and cell viability |
title_sort | ran-dependent docking of importin-β to ranbp2/nup358 filaments is essential for protein import and cell viability |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3160583/ https://www.ncbi.nlm.nih.gov/pubmed/21859863 http://dx.doi.org/10.1083/jcb.201102018 |
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