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Septin filaments exhibit a dynamic, paired organization that is conserved from yeast to mammals
The septins are conserved, GTP-binding proteins important for cytokinesis, membrane compartmentalization, and exocytosis. However, it is unknown how septins are arranged within higher-order structures in cells. To determine the organization of septins in live cells, we developed a polarized fluoresc...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Rockefeller University Press
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3115802/ https://www.ncbi.nlm.nih.gov/pubmed/21670216 http://dx.doi.org/10.1083/jcb.201012143 |
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author | DeMay, Bradley S. Bai, Xiaobo Howard, Louisa Occhipinti, Patricia Meseroll, Rebecca A. Spiliotis, Elias T. Oldenbourg, Rudolf Gladfelter, Amy S. |
author_facet | DeMay, Bradley S. Bai, Xiaobo Howard, Louisa Occhipinti, Patricia Meseroll, Rebecca A. Spiliotis, Elias T. Oldenbourg, Rudolf Gladfelter, Amy S. |
author_sort | DeMay, Bradley S. |
collection | PubMed |
description | The septins are conserved, GTP-binding proteins important for cytokinesis, membrane compartmentalization, and exocytosis. However, it is unknown how septins are arranged within higher-order structures in cells. To determine the organization of septins in live cells, we developed a polarized fluorescence microscopy system to monitor the orientation of GFP dipole moments with high spatial and temporal resolution. When GFP was fused to septins, the arrangement of GFP dipoles reflected the underlying septin organization. We demonstrated in a filamentous fungus, a budding yeast, and a mammalian epithelial cell line that septin proteins were organized in an identical highly ordered fashion. Fluorescence anisotropy measurements indicated that septin filaments organized into pairs within live cells, just as has been observed in vitro. Additional support for the formation of pairs came from the observation of paired filaments at the cortex of cells using electron microscopy. Furthermore, we found that highly ordered septin structures exchanged subunits and rapidly rearranged. We conclude that septins assemble into dynamic, paired filaments in vivo and that this organization is conserved from yeast to mammals. |
format | Online Article Text |
id | pubmed-3115802 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | The Rockefeller University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-31158022011-12-13 Septin filaments exhibit a dynamic, paired organization that is conserved from yeast to mammals DeMay, Bradley S. Bai, Xiaobo Howard, Louisa Occhipinti, Patricia Meseroll, Rebecca A. Spiliotis, Elias T. Oldenbourg, Rudolf Gladfelter, Amy S. J Cell Biol Research Articles The septins are conserved, GTP-binding proteins important for cytokinesis, membrane compartmentalization, and exocytosis. However, it is unknown how septins are arranged within higher-order structures in cells. To determine the organization of septins in live cells, we developed a polarized fluorescence microscopy system to monitor the orientation of GFP dipole moments with high spatial and temporal resolution. When GFP was fused to septins, the arrangement of GFP dipoles reflected the underlying septin organization. We demonstrated in a filamentous fungus, a budding yeast, and a mammalian epithelial cell line that septin proteins were organized in an identical highly ordered fashion. Fluorescence anisotropy measurements indicated that septin filaments organized into pairs within live cells, just as has been observed in vitro. Additional support for the formation of pairs came from the observation of paired filaments at the cortex of cells using electron microscopy. Furthermore, we found that highly ordered septin structures exchanged subunits and rapidly rearranged. We conclude that septins assemble into dynamic, paired filaments in vivo and that this organization is conserved from yeast to mammals. The Rockefeller University Press 2011-06-13 /pmc/articles/PMC3115802/ /pubmed/21670216 http://dx.doi.org/10.1083/jcb.201012143 Text en © 2011 DeMay et al. 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 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/). |
spellingShingle | Research Articles DeMay, Bradley S. Bai, Xiaobo Howard, Louisa Occhipinti, Patricia Meseroll, Rebecca A. Spiliotis, Elias T. Oldenbourg, Rudolf Gladfelter, Amy S. Septin filaments exhibit a dynamic, paired organization that is conserved from yeast to mammals |
title | Septin filaments exhibit a dynamic, paired organization that is conserved from yeast to mammals |
title_full | Septin filaments exhibit a dynamic, paired organization that is conserved from yeast to mammals |
title_fullStr | Septin filaments exhibit a dynamic, paired organization that is conserved from yeast to mammals |
title_full_unstemmed | Septin filaments exhibit a dynamic, paired organization that is conserved from yeast to mammals |
title_short | Septin filaments exhibit a dynamic, paired organization that is conserved from yeast to mammals |
title_sort | septin filaments exhibit a dynamic, paired organization that is conserved from yeast to mammals |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3115802/ https://www.ncbi.nlm.nih.gov/pubmed/21670216 http://dx.doi.org/10.1083/jcb.201012143 |
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