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Visualizing the dynamic coupling of claudin strands to the actin cytoskeleton through ZO-1
The organization and integrity of epithelial tight junctions depend on interactions between claudins, ZO scaffolding proteins, and the cytoskeleton. However, although binding between claudins and ZO-1/2/3 and between ZO-1/2/3 and numerous cytoskeletal proteins has been demonstrated in vitro, fluores...
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The American Society for Cell Biology
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5305259/ https://www.ncbi.nlm.nih.gov/pubmed/27974639 http://dx.doi.org/10.1091/mbc.E16-10-0698 |
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author | Van Itallie, Christina M. Tietgens, Amber Jean Anderson, James M. |
author_facet | Van Itallie, Christina M. Tietgens, Amber Jean Anderson, James M. |
author_sort | Van Itallie, Christina M. |
collection | PubMed |
description | The organization and integrity of epithelial tight junctions depend on interactions between claudins, ZO scaffolding proteins, and the cytoskeleton. However, although binding between claudins and ZO-1/2/3 and between ZO-1/2/3 and numerous cytoskeletal proteins has been demonstrated in vitro, fluorescence recovery after photobleaching analysis suggests interactions in vivo are likely highly dynamic. Here we use superresolution live-cell imaging in a model fibroblast system to examine relationships between claudins, ZO-1, occludin, and actin. We find that GFP claudins make easily visualized dynamic strand patches between two fibroblasts; strand dynamics is constrained by ZO-1 binding. Claudin association with actin is also dependent on ZO-1, but colocalization demonstrates intermittent rather than continuous association between claudin, ZO-1, and actin. Independent of interaction with ZO-1 or actin, claudin strands break and reanneal; pulse-chase-pulse analysis using SNAP-tagged claudins showed preferential incorporation of newly synthesized claudins into break sites. Although claudin strand behavior in fibroblasts may not fully recapitulate that of epithelial tight junction strands, this is the first direct demonstration of the ability of ZO-1 to stabilize claudin strands. We speculate that intermittent tethering of claudins to actin may allow for accommodation of the paracellular seal to physiological or pathological alterations in cell shape or movement. |
format | Online Article Text |
id | pubmed-5305259 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-53052592017-04-30 Visualizing the dynamic coupling of claudin strands to the actin cytoskeleton through ZO-1 Van Itallie, Christina M. Tietgens, Amber Jean Anderson, James M. Mol Biol Cell Articles The organization and integrity of epithelial tight junctions depend on interactions between claudins, ZO scaffolding proteins, and the cytoskeleton. However, although binding between claudins and ZO-1/2/3 and between ZO-1/2/3 and numerous cytoskeletal proteins has been demonstrated in vitro, fluorescence recovery after photobleaching analysis suggests interactions in vivo are likely highly dynamic. Here we use superresolution live-cell imaging in a model fibroblast system to examine relationships between claudins, ZO-1, occludin, and actin. We find that GFP claudins make easily visualized dynamic strand patches between two fibroblasts; strand dynamics is constrained by ZO-1 binding. Claudin association with actin is also dependent on ZO-1, but colocalization demonstrates intermittent rather than continuous association between claudin, ZO-1, and actin. Independent of interaction with ZO-1 or actin, claudin strands break and reanneal; pulse-chase-pulse analysis using SNAP-tagged claudins showed preferential incorporation of newly synthesized claudins into break sites. Although claudin strand behavior in fibroblasts may not fully recapitulate that of epithelial tight junction strands, this is the first direct demonstration of the ability of ZO-1 to stabilize claudin strands. We speculate that intermittent tethering of claudins to actin may allow for accommodation of the paracellular seal to physiological or pathological alterations in cell shape or movement. The American Society for Cell Biology 2017-02-15 /pmc/articles/PMC5305259/ /pubmed/27974639 http://dx.doi.org/10.1091/mbc.E16-10-0698 Text en © 2017 Van Itallie 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 | Articles Van Itallie, Christina M. Tietgens, Amber Jean Anderson, James M. Visualizing the dynamic coupling of claudin strands to the actin cytoskeleton through ZO-1 |
title | Visualizing the dynamic coupling of claudin strands to the actin cytoskeleton through ZO-1 |
title_full | Visualizing the dynamic coupling of claudin strands to the actin cytoskeleton through ZO-1 |
title_fullStr | Visualizing the dynamic coupling of claudin strands to the actin cytoskeleton through ZO-1 |
title_full_unstemmed | Visualizing the dynamic coupling of claudin strands to the actin cytoskeleton through ZO-1 |
title_short | Visualizing the dynamic coupling of claudin strands to the actin cytoskeleton through ZO-1 |
title_sort | visualizing the dynamic coupling of claudin strands to the actin cytoskeleton through zo-1 |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5305259/ https://www.ncbi.nlm.nih.gov/pubmed/27974639 http://dx.doi.org/10.1091/mbc.E16-10-0698 |
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