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Effects of Plectin Depletion on Keratin Network Dynamics and Organization
The keratin intermediate filament cytoskeleton protects epithelial cells against various types of stress and is involved in fundamental cellular processes such as signaling, differentiation and organelle trafficking. These functions rely on the cell type-specific arrangement and plasticity of the ke...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4805305/ https://www.ncbi.nlm.nih.gov/pubmed/27007410 http://dx.doi.org/10.1371/journal.pone.0149106 |
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author | Moch, Marcin Windoffer, Reinhard Schwarz, Nicole Pohl, Raphaela Omenzetter, Andreas Schnakenberg, Uwe Herb, Fabian Chaisaowong, Kraisorn Merhof, Dorit Ramms, Lena Fabris, Gloria Hoffmann, Bernd Merkel, Rudolf Leube, Rudolf E. |
author_facet | Moch, Marcin Windoffer, Reinhard Schwarz, Nicole Pohl, Raphaela Omenzetter, Andreas Schnakenberg, Uwe Herb, Fabian Chaisaowong, Kraisorn Merhof, Dorit Ramms, Lena Fabris, Gloria Hoffmann, Bernd Merkel, Rudolf Leube, Rudolf E. |
author_sort | Moch, Marcin |
collection | PubMed |
description | The keratin intermediate filament cytoskeleton protects epithelial cells against various types of stress and is involved in fundamental cellular processes such as signaling, differentiation and organelle trafficking. These functions rely on the cell type-specific arrangement and plasticity of the keratin system. It has been suggested that these properties are regulated by a complex cycle of assembly and disassembly. The exact mechanisms responsible for the underlying molecular processes, however, have not been clarified. Accumulating evidence implicates the cytolinker plectin in various aspects of the keratin cycle, i.e., by acting as a stabilizing anchor at hemidesmosomal adhesion sites and the nucleus, by affecting keratin bundling and branching and by linkage of keratins to actin filament and microtubule dynamics. In the present study we tested these hypotheses. To this end, plectin was downregulated by shRNA in vulvar carcinoma-derived A431 cells. As expected, integrin β4- and BPAG-1-positive hemidesmosomal structures were strongly reduced and cytosolic actin stress fibers were increased. In addition, integrins α3 and β1 were reduced. The experiments furthermore showed that loss of plectin led to a reduction in keratin filament branch length but did not alter overall mechanical properties as assessed by indentation analyses using atomic force microscopy and by displacement analyses of cytoplasmic superparamagnetic beads using magnetic tweezers. An increase in keratin movement was observed in plectin-depleted cells as was the case in control cells lacking hemidesmosome-like structures. Yet, keratin turnover was not significantly affected. We conclude that plectin alone is not needed for keratin assembly and disassembly and that other mechanisms exist to guarantee proper keratin cycling under steady state conditions in cultured single cells. |
format | Online Article Text |
id | pubmed-4805305 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-48053052016-03-25 Effects of Plectin Depletion on Keratin Network Dynamics and Organization Moch, Marcin Windoffer, Reinhard Schwarz, Nicole Pohl, Raphaela Omenzetter, Andreas Schnakenberg, Uwe Herb, Fabian Chaisaowong, Kraisorn Merhof, Dorit Ramms, Lena Fabris, Gloria Hoffmann, Bernd Merkel, Rudolf Leube, Rudolf E. PLoS One Research Article The keratin intermediate filament cytoskeleton protects epithelial cells against various types of stress and is involved in fundamental cellular processes such as signaling, differentiation and organelle trafficking. These functions rely on the cell type-specific arrangement and plasticity of the keratin system. It has been suggested that these properties are regulated by a complex cycle of assembly and disassembly. The exact mechanisms responsible for the underlying molecular processes, however, have not been clarified. Accumulating evidence implicates the cytolinker plectin in various aspects of the keratin cycle, i.e., by acting as a stabilizing anchor at hemidesmosomal adhesion sites and the nucleus, by affecting keratin bundling and branching and by linkage of keratins to actin filament and microtubule dynamics. In the present study we tested these hypotheses. To this end, plectin was downregulated by shRNA in vulvar carcinoma-derived A431 cells. As expected, integrin β4- and BPAG-1-positive hemidesmosomal structures were strongly reduced and cytosolic actin stress fibers were increased. In addition, integrins α3 and β1 were reduced. The experiments furthermore showed that loss of plectin led to a reduction in keratin filament branch length but did not alter overall mechanical properties as assessed by indentation analyses using atomic force microscopy and by displacement analyses of cytoplasmic superparamagnetic beads using magnetic tweezers. An increase in keratin movement was observed in plectin-depleted cells as was the case in control cells lacking hemidesmosome-like structures. Yet, keratin turnover was not significantly affected. We conclude that plectin alone is not needed for keratin assembly and disassembly and that other mechanisms exist to guarantee proper keratin cycling under steady state conditions in cultured single cells. Public Library of Science 2016-03-23 /pmc/articles/PMC4805305/ /pubmed/27007410 http://dx.doi.org/10.1371/journal.pone.0149106 Text en © 2016 Moch et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Moch, Marcin Windoffer, Reinhard Schwarz, Nicole Pohl, Raphaela Omenzetter, Andreas Schnakenberg, Uwe Herb, Fabian Chaisaowong, Kraisorn Merhof, Dorit Ramms, Lena Fabris, Gloria Hoffmann, Bernd Merkel, Rudolf Leube, Rudolf E. Effects of Plectin Depletion on Keratin Network Dynamics and Organization |
title | Effects of Plectin Depletion on Keratin Network Dynamics and Organization |
title_full | Effects of Plectin Depletion on Keratin Network Dynamics and Organization |
title_fullStr | Effects of Plectin Depletion on Keratin Network Dynamics and Organization |
title_full_unstemmed | Effects of Plectin Depletion on Keratin Network Dynamics and Organization |
title_short | Effects of Plectin Depletion on Keratin Network Dynamics and Organization |
title_sort | effects of plectin depletion on keratin network dynamics and organization |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4805305/ https://www.ncbi.nlm.nih.gov/pubmed/27007410 http://dx.doi.org/10.1371/journal.pone.0149106 |
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