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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...

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Autores principales: Hamada, Masakazu, Haeger, Anna, Jeganathan, Karthik B., van Ree, Janine H., Malureanu, Liviu, Wälde, Sarah, Joseph, Jomon, Kehlenbach, Ralph H., van Deursen, Jan M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Rockefeller University Press 2011
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.
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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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