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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...

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Autores principales: Laudermilch, Ethan, Tsai, Pei-Ling, Graham, Morven, Turner, Elizabeth, Zhao, Chenguang, Schlieker, Christian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2016
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.
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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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