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NuMA localization, stability, and function in spindle orientation involve 4.1 and Cdk1 interactions
The epidermis is a multilayered epithelium that requires asymmetric divisions for stratification. A conserved cortical protein complex, including LGN, nuclear mitotic apparatus (NuMA), and dynein/dynactin, plays a key role in establishing proper spindle orientation during asymmetric divisions. The r...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The American Society for Cell Biology
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3842992/ https://www.ncbi.nlm.nih.gov/pubmed/24109598 http://dx.doi.org/10.1091/mbc.E13-05-0277 |
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author | Seldin, Lindsey Poulson, Nicholas D. Foote, Henry P. Lechler, Terry |
author_facet | Seldin, Lindsey Poulson, Nicholas D. Foote, Henry P. Lechler, Terry |
author_sort | Seldin, Lindsey |
collection | PubMed |
description | The epidermis is a multilayered epithelium that requires asymmetric divisions for stratification. A conserved cortical protein complex, including LGN, nuclear mitotic apparatus (NuMA), and dynein/dynactin, plays a key role in establishing proper spindle orientation during asymmetric divisions. The requirements for the cortical recruitment of these proteins, however, remain unclear. In this work, we show that NuMA is required to recruit dynactin to the cell cortex of keratinocytes. NuMA's cortical recruitment requires LGN; however, LGN interactions are not sufficient for this localization. Using fluorescence recovery after photobleaching, we find that the 4.1-binding domain of NuMA is important for stabilizing its interaction with the cell cortex. This is functionally important, as loss of 4.1/NuMA interaction results in spindle orientation defects, using two distinct assays. Furthermore, we observe an increase in cortical NuMA localization as cells enter anaphase. Inhibition of Cdk1 or mutation of a single residue in NuMA mimics this effect. NuMA's anaphase localization is independent of LGN and 4.1 interactions, revealing two distinct mechanisms responsible for NuMA cortical recruitment at different stages of mitosis. This work highlights the complexity of NuMA localization and reveals the importance of NuMA cortical stability for productive force generation during spindle orientation. |
format | Online Article Text |
id | pubmed-3842992 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-38429922014-02-16 NuMA localization, stability, and function in spindle orientation involve 4.1 and Cdk1 interactions Seldin, Lindsey Poulson, Nicholas D. Foote, Henry P. Lechler, Terry Mol Biol Cell Articles The epidermis is a multilayered epithelium that requires asymmetric divisions for stratification. A conserved cortical protein complex, including LGN, nuclear mitotic apparatus (NuMA), and dynein/dynactin, plays a key role in establishing proper spindle orientation during asymmetric divisions. The requirements for the cortical recruitment of these proteins, however, remain unclear. In this work, we show that NuMA is required to recruit dynactin to the cell cortex of keratinocytes. NuMA's cortical recruitment requires LGN; however, LGN interactions are not sufficient for this localization. Using fluorescence recovery after photobleaching, we find that the 4.1-binding domain of NuMA is important for stabilizing its interaction with the cell cortex. This is functionally important, as loss of 4.1/NuMA interaction results in spindle orientation defects, using two distinct assays. Furthermore, we observe an increase in cortical NuMA localization as cells enter anaphase. Inhibition of Cdk1 or mutation of a single residue in NuMA mimics this effect. NuMA's anaphase localization is independent of LGN and 4.1 interactions, revealing two distinct mechanisms responsible for NuMA cortical recruitment at different stages of mitosis. This work highlights the complexity of NuMA localization and reveals the importance of NuMA cortical stability for productive force generation during spindle orientation. The American Society for Cell Biology 2013-12-01 /pmc/articles/PMC3842992/ /pubmed/24109598 http://dx.doi.org/10.1091/mbc.E13-05-0277 Text en © 2013 Seldin 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 of Cell Biology. |
spellingShingle | Articles Seldin, Lindsey Poulson, Nicholas D. Foote, Henry P. Lechler, Terry NuMA localization, stability, and function in spindle orientation involve 4.1 and Cdk1 interactions |
title | NuMA localization, stability, and function in spindle orientation involve 4.1 and Cdk1 interactions |
title_full | NuMA localization, stability, and function in spindle orientation involve 4.1 and Cdk1 interactions |
title_fullStr | NuMA localization, stability, and function in spindle orientation involve 4.1 and Cdk1 interactions |
title_full_unstemmed | NuMA localization, stability, and function in spindle orientation involve 4.1 and Cdk1 interactions |
title_short | NuMA localization, stability, and function in spindle orientation involve 4.1 and Cdk1 interactions |
title_sort | numa localization, stability, and function in spindle orientation involve 4.1 and cdk1 interactions |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3842992/ https://www.ncbi.nlm.nih.gov/pubmed/24109598 http://dx.doi.org/10.1091/mbc.E13-05-0277 |
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