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Imaging within single NPCs reveals NXF1’s role in mRNA export on the cytoplasmic side of the pore

Translocation of mRNA through the nuclear pore complex (NPC) requires interactions with different NPC regions. To determine the interactions that are crucial for effective mRNA export in living cells, we examined mRNA export within individual pores by applying various types of mRNA export blocks tha...

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Autores principales: Ben-Yishay, Rakefet, Mor, Amir, Shraga, Amit, Ashkenazy-Titelman, Asaf, Kinor, Noa, Schwed-Gross, Avital, Jacob, Avi, Kozer, Noga, Kumar, Pramod, Garini, Yuval, Shav-Tal, Yaron
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Rockefeller University Press 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6719458/
https://www.ncbi.nlm.nih.gov/pubmed/31375530
http://dx.doi.org/10.1083/jcb.201901127
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author Ben-Yishay, Rakefet
Mor, Amir
Shraga, Amit
Ashkenazy-Titelman, Asaf
Kinor, Noa
Schwed-Gross, Avital
Jacob, Avi
Kozer, Noga
Kumar, Pramod
Garini, Yuval
Shav-Tal, Yaron
author_facet Ben-Yishay, Rakefet
Mor, Amir
Shraga, Amit
Ashkenazy-Titelman, Asaf
Kinor, Noa
Schwed-Gross, Avital
Jacob, Avi
Kozer, Noga
Kumar, Pramod
Garini, Yuval
Shav-Tal, Yaron
author_sort Ben-Yishay, Rakefet
collection PubMed
description Translocation of mRNA through the nuclear pore complex (NPC) requires interactions with different NPC regions. To determine the interactions that are crucial for effective mRNA export in living cells, we examined mRNA export within individual pores by applying various types of mRNA export blocks that stalled mRNPs at different stages of transition. Focusing on the major mRNA export factor NXF1, we found that initial mRNP binding to the NPC did not require NXF1 in the NPC, whereas release into the cytoplasm did. NXF1 localization in the NPC did not require RNA or RNA binding. Superresolution microscopy showed that NXF1 consistently occupied positions on the cytoplasmic side of the NPC. Interactions with specific nucleoporins were pinpointed using FLIM-FRET for measuring protein–protein interactions inside single NPCs, showing that Dbp5 helicase activity of mRNA release is conserved in yeast and humans. Altogether, we find that specific interactions on the cytoplasmic side of the NPC are fundamental for the directional flow of mRNA export.
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spelling pubmed-67194582020-03-02 Imaging within single NPCs reveals NXF1’s role in mRNA export on the cytoplasmic side of the pore Ben-Yishay, Rakefet Mor, Amir Shraga, Amit Ashkenazy-Titelman, Asaf Kinor, Noa Schwed-Gross, Avital Jacob, Avi Kozer, Noga Kumar, Pramod Garini, Yuval Shav-Tal, Yaron J Cell Biol Research Articles Translocation of mRNA through the nuclear pore complex (NPC) requires interactions with different NPC regions. To determine the interactions that are crucial for effective mRNA export in living cells, we examined mRNA export within individual pores by applying various types of mRNA export blocks that stalled mRNPs at different stages of transition. Focusing on the major mRNA export factor NXF1, we found that initial mRNP binding to the NPC did not require NXF1 in the NPC, whereas release into the cytoplasm did. NXF1 localization in the NPC did not require RNA or RNA binding. Superresolution microscopy showed that NXF1 consistently occupied positions on the cytoplasmic side of the NPC. Interactions with specific nucleoporins were pinpointed using FLIM-FRET for measuring protein–protein interactions inside single NPCs, showing that Dbp5 helicase activity of mRNA release is conserved in yeast and humans. Altogether, we find that specific interactions on the cytoplasmic side of the NPC are fundamental for the directional flow of mRNA export. Rockefeller University Press 2019-09-02 2019-08-02 /pmc/articles/PMC6719458/ /pubmed/31375530 http://dx.doi.org/10.1083/jcb.201901127 Text en © 2019 Ben-Yishay et al. http://www.rupress.org/terms/https://creativecommons.org/licenses/by-nc-sa/4.0/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 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/).
spellingShingle Research Articles
Ben-Yishay, Rakefet
Mor, Amir
Shraga, Amit
Ashkenazy-Titelman, Asaf
Kinor, Noa
Schwed-Gross, Avital
Jacob, Avi
Kozer, Noga
Kumar, Pramod
Garini, Yuval
Shav-Tal, Yaron
Imaging within single NPCs reveals NXF1’s role in mRNA export on the cytoplasmic side of the pore
title Imaging within single NPCs reveals NXF1’s role in mRNA export on the cytoplasmic side of the pore
title_full Imaging within single NPCs reveals NXF1’s role in mRNA export on the cytoplasmic side of the pore
title_fullStr Imaging within single NPCs reveals NXF1’s role in mRNA export on the cytoplasmic side of the pore
title_full_unstemmed Imaging within single NPCs reveals NXF1’s role in mRNA export on the cytoplasmic side of the pore
title_short Imaging within single NPCs reveals NXF1’s role in mRNA export on the cytoplasmic side of the pore
title_sort imaging within single npcs reveals nxf1’s role in mrna export on the cytoplasmic side of the pore
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6719458/
https://www.ncbi.nlm.nih.gov/pubmed/31375530
http://dx.doi.org/10.1083/jcb.201901127
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