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Reticulon and CLIMP-63 regulate nanodomain organization of peripheral ER tubules
The endoplasmic reticulum (ER) is an expansive, membrane-enclosed organelle composed of smooth peripheral tubules and rough, ribosome-studded central ER sheets whose morphology is determined, in part, by the ER-shaping proteins, reticulon (RTN) and cytoskeleton-linking membrane protein 63 (CLIMP-63)...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6742417/ https://www.ncbi.nlm.nih.gov/pubmed/31469817 http://dx.doi.org/10.1371/journal.pbio.3000355 |
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author | Gao, Guang Zhu, Chengjia Liu, Emma Nabi, Ivan R. |
author_facet | Gao, Guang Zhu, Chengjia Liu, Emma Nabi, Ivan R. |
author_sort | Gao, Guang |
collection | PubMed |
description | The endoplasmic reticulum (ER) is an expansive, membrane-enclosed organelle composed of smooth peripheral tubules and rough, ribosome-studded central ER sheets whose morphology is determined, in part, by the ER-shaping proteins, reticulon (RTN) and cytoskeleton-linking membrane protein 63 (CLIMP-63), respectively. Here, stimulated emission depletion (STED) super-resolution microscopy shows that reticulon4a (RTN4a) and CLIMP-63 also regulate the organization and dynamics of peripheral ER tubule nanodomains. STED imaging shows that lumenal ER monomeric oxidizing environment-optimized green fluorescent protein (ERmoxGFP), membrane Sec61βGFP, knock-in calreticulin-GFP, and antibody-labeled ER-resident proteins calnexin and derlin-1 are all localized to periodic puncta along the length of peripheral ER tubules that are not readily observable by diffraction limited confocal microscopy. RTN4a segregates away from and restricts lumenal blob length, while CLIMP-63 associates with and increases lumenal blob length. RTN4a and CLIMP-63 also regulate the nanodomain distribution of ER-resident proteins, being required for the preferential segregation of calnexin and derlin-1 puncta away from lumenal ERmoxGFP blobs. High-speed (40 ms/frame) live cell STED imaging shows that RTN4a and CLIMP-63 regulate dynamic nanoscale lumenal compartmentalization along peripheral ER tubules. RTN4a enhances and CLIMP-63 disrupts the local accumulation of lumenal ERmoxGFP at spatially defined sites along ER tubules. The ER-shaping proteins RTN and CLIMP-63 therefore regulate lumenal ER nanodomain heterogeneity, interaction with ER-resident proteins, and dynamics in peripheral ER tubules. |
format | Online Article Text |
id | pubmed-6742417 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-67424172019-09-20 Reticulon and CLIMP-63 regulate nanodomain organization of peripheral ER tubules Gao, Guang Zhu, Chengjia Liu, Emma Nabi, Ivan R. PLoS Biol Research Article The endoplasmic reticulum (ER) is an expansive, membrane-enclosed organelle composed of smooth peripheral tubules and rough, ribosome-studded central ER sheets whose morphology is determined, in part, by the ER-shaping proteins, reticulon (RTN) and cytoskeleton-linking membrane protein 63 (CLIMP-63), respectively. Here, stimulated emission depletion (STED) super-resolution microscopy shows that reticulon4a (RTN4a) and CLIMP-63 also regulate the organization and dynamics of peripheral ER tubule nanodomains. STED imaging shows that lumenal ER monomeric oxidizing environment-optimized green fluorescent protein (ERmoxGFP), membrane Sec61βGFP, knock-in calreticulin-GFP, and antibody-labeled ER-resident proteins calnexin and derlin-1 are all localized to periodic puncta along the length of peripheral ER tubules that are not readily observable by diffraction limited confocal microscopy. RTN4a segregates away from and restricts lumenal blob length, while CLIMP-63 associates with and increases lumenal blob length. RTN4a and CLIMP-63 also regulate the nanodomain distribution of ER-resident proteins, being required for the preferential segregation of calnexin and derlin-1 puncta away from lumenal ERmoxGFP blobs. High-speed (40 ms/frame) live cell STED imaging shows that RTN4a and CLIMP-63 regulate dynamic nanoscale lumenal compartmentalization along peripheral ER tubules. RTN4a enhances and CLIMP-63 disrupts the local accumulation of lumenal ERmoxGFP at spatially defined sites along ER tubules. The ER-shaping proteins RTN and CLIMP-63 therefore regulate lumenal ER nanodomain heterogeneity, interaction with ER-resident proteins, and dynamics in peripheral ER tubules. Public Library of Science 2019-08-30 /pmc/articles/PMC6742417/ /pubmed/31469817 http://dx.doi.org/10.1371/journal.pbio.3000355 Text en © 2019 Gao et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Gao, Guang Zhu, Chengjia Liu, Emma Nabi, Ivan R. Reticulon and CLIMP-63 regulate nanodomain organization of peripheral ER tubules |
title | Reticulon and CLIMP-63 regulate nanodomain organization of peripheral ER tubules |
title_full | Reticulon and CLIMP-63 regulate nanodomain organization of peripheral ER tubules |
title_fullStr | Reticulon and CLIMP-63 regulate nanodomain organization of peripheral ER tubules |
title_full_unstemmed | Reticulon and CLIMP-63 regulate nanodomain organization of peripheral ER tubules |
title_short | Reticulon and CLIMP-63 regulate nanodomain organization of peripheral ER tubules |
title_sort | reticulon and climp-63 regulate nanodomain organization of peripheral er tubules |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6742417/ https://www.ncbi.nlm.nih.gov/pubmed/31469817 http://dx.doi.org/10.1371/journal.pbio.3000355 |
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