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Dissecting Torsin/cofactor function at the nuclear envelope: a genetic study
The human genome encodes four Torsin ATPases, the functions of which are poorly understood. In this study, we use CRISPR/Cas9 engineering to delete all four Torsin ATPases individually and in combination. Using nuclear envelope (NE) blebbing as a phenotypic measure, we establish a direct correlation...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The American Society for Cell Biology
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5156537/ https://www.ncbi.nlm.nih.gov/pubmed/27798237 http://dx.doi.org/10.1091/mbc.E16-07-0511 |
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author | Laudermilch, Ethan Tsai, Pei-Ling Graham, Morven Turner, Elizabeth Zhao, Chenguang Schlieker, Christian |
author_facet | Laudermilch, Ethan Tsai, Pei-Ling Graham, Morven Turner, Elizabeth Zhao, Chenguang Schlieker, Christian |
author_sort | Laudermilch, Ethan |
collection | PubMed |
description | The human genome encodes four Torsin ATPases, the functions of which are poorly understood. In this study, we use CRISPR/Cas9 engineering to delete all four Torsin ATPases individually and in combination. Using nuclear envelope (NE) blebbing as a phenotypic measure, we establish a direct correlation between the number of inactivated Torsin alleles and the occurrence of omega-shaped herniations within the lumen of the NE. A similar, although not identical, redundancy is observed for LAP1 and LULL1, which serve as regulatory cofactors for a subset of Torsin ATPases. Unexpectedly, deletion of Tor2A in a TorA/B/3A-deficient background results in a stark increase of bleb formation, even though Tor2A does not respond to LAP1/LULL1 stimulation. The robustness of the observed phenotype in Torsin-deficient cells enables a structural analysis via electron microscopy tomography and a compositional analysis via immunogold labeling. Ubiquitin and nucleoporins were identified as distinctively localizing components of the omega-shaped bleb structure. These findings suggest a functional link between the Torsin/cofactor system and NE/nuclear pore complex biogenesis or homeostasis and establish a Torsin-deficient cell line as a valuable experimental platform with which to decipher Torsin function. |
format | Online Article Text |
id | pubmed-5156537 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-51565372017-03-02 Dissecting Torsin/cofactor function at the nuclear envelope: a genetic study Laudermilch, Ethan Tsai, Pei-Ling Graham, Morven Turner, Elizabeth Zhao, Chenguang Schlieker, Christian Mol Biol Cell Brief Reports The human genome encodes four Torsin ATPases, the functions of which are poorly understood. In this study, we use CRISPR/Cas9 engineering to delete all four Torsin ATPases individually and in combination. Using nuclear envelope (NE) blebbing as a phenotypic measure, we establish a direct correlation between the number of inactivated Torsin alleles and the occurrence of omega-shaped herniations within the lumen of the NE. A similar, although not identical, redundancy is observed for LAP1 and LULL1, which serve as regulatory cofactors for a subset of Torsin ATPases. Unexpectedly, deletion of Tor2A in a TorA/B/3A-deficient background results in a stark increase of bleb formation, even though Tor2A does not respond to LAP1/LULL1 stimulation. The robustness of the observed phenotype in Torsin-deficient cells enables a structural analysis via electron microscopy tomography and a compositional analysis via immunogold labeling. Ubiquitin and nucleoporins were identified as distinctively localizing components of the omega-shaped bleb structure. These findings suggest a functional link between the Torsin/cofactor system and NE/nuclear pore complex biogenesis or homeostasis and establish a Torsin-deficient cell line as a valuable experimental platform with which to decipher Torsin function. The American Society for Cell Biology 2016-12-15 /pmc/articles/PMC5156537/ /pubmed/27798237 http://dx.doi.org/10.1091/mbc.E16-07-0511 Text en © 2016 Laudermilch et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology. |
spellingShingle | Brief Reports Laudermilch, Ethan Tsai, Pei-Ling Graham, Morven Turner, Elizabeth Zhao, Chenguang Schlieker, Christian Dissecting Torsin/cofactor function at the nuclear envelope: a genetic study |
title | Dissecting Torsin/cofactor function at the nuclear envelope: a genetic study |
title_full | Dissecting Torsin/cofactor function at the nuclear envelope: a genetic study |
title_fullStr | Dissecting Torsin/cofactor function at the nuclear envelope: a genetic study |
title_full_unstemmed | Dissecting Torsin/cofactor function at the nuclear envelope: a genetic study |
title_short | Dissecting Torsin/cofactor function at the nuclear envelope: a genetic study |
title_sort | dissecting torsin/cofactor function at the nuclear envelope: a genetic study |
topic | Brief Reports |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5156537/ https://www.ncbi.nlm.nih.gov/pubmed/27798237 http://dx.doi.org/10.1091/mbc.E16-07-0511 |
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