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The elusive actin cytoskeleton of a green alga expressing both conventional and divergent actins
The green alga Chlamydomonas reinhardtii is a leading model system to study photosynthesis, cilia, and the generation of biological products. The cytoskeleton plays important roles in all of these cellular processes, but to date, the filamentous actin network within Chlamydomonas has remained elusiv...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The American Society for Cell Biology
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6789165/ https://www.ncbi.nlm.nih.gov/pubmed/31532705 http://dx.doi.org/10.1091/mbc.E19-03-0141 |
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author | Craig, Evan W. Mueller, David M. Bigge, Brae M. Schaffer, Miroslava Engel, Benjamin D. Avasthi, Prachee |
author_facet | Craig, Evan W. Mueller, David M. Bigge, Brae M. Schaffer, Miroslava Engel, Benjamin D. Avasthi, Prachee |
author_sort | Craig, Evan W. |
collection | PubMed |
description | The green alga Chlamydomonas reinhardtii is a leading model system to study photosynthesis, cilia, and the generation of biological products. The cytoskeleton plays important roles in all of these cellular processes, but to date, the filamentous actin network within Chlamydomonas has remained elusive. By optimizing labeling conditions, we can now visualize distinct linear actin filaments at the posterior of the nucleus in both live and fixed vegetative cells. Using in situ cryo-electron tomography, we confirmed this localization by directly imaging actin filaments within the native cellular environment. The fluorescently labeled structures are sensitive to the depolymerizing agent latrunculin B (Lat B), demonstrating the specificity of our optimized labeling method. Interestingly, Lat B treatment resulted in the formation of a transient ring-like filamentous actin structure around the nucleus. The assembly of this perinuclear ring is dependent upon a second actin isoform, NAP1, which is strongly up-regulated upon Lat B treatment and is insensitive to Lat B–induced depolymerization. Our study combines orthogonal strategies to provide the first detailed visual characterization of filamentous actins in Chlamydomonas, allowing insights into the coordinated functions of two actin isoforms expressed within the same cell. |
format | Online Article Text |
id | pubmed-6789165 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-67891652019-12-30 The elusive actin cytoskeleton of a green alga expressing both conventional and divergent actins Craig, Evan W. Mueller, David M. Bigge, Brae M. Schaffer, Miroslava Engel, Benjamin D. Avasthi, Prachee Mol Biol Cell Article The green alga Chlamydomonas reinhardtii is a leading model system to study photosynthesis, cilia, and the generation of biological products. The cytoskeleton plays important roles in all of these cellular processes, but to date, the filamentous actin network within Chlamydomonas has remained elusive. By optimizing labeling conditions, we can now visualize distinct linear actin filaments at the posterior of the nucleus in both live and fixed vegetative cells. Using in situ cryo-electron tomography, we confirmed this localization by directly imaging actin filaments within the native cellular environment. The fluorescently labeled structures are sensitive to the depolymerizing agent latrunculin B (Lat B), demonstrating the specificity of our optimized labeling method. Interestingly, Lat B treatment resulted in the formation of a transient ring-like filamentous actin structure around the nucleus. The assembly of this perinuclear ring is dependent upon a second actin isoform, NAP1, which is strongly up-regulated upon Lat B treatment and is insensitive to Lat B–induced depolymerization. Our study combines orthogonal strategies to provide the first detailed visual characterization of filamentous actins in Chlamydomonas, allowing insights into the coordinated functions of two actin isoforms expressed within the same cell. The American Society for Cell Biology 2019-10-15 /pmc/articles/PMC6789165/ /pubmed/31532705 http://dx.doi.org/10.1091/mbc.E19-03-0141 Text en © 2019 Craig et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology. http://creativecommons.org/licenses/by-nc-sa/3.0 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. |
spellingShingle | Article Craig, Evan W. Mueller, David M. Bigge, Brae M. Schaffer, Miroslava Engel, Benjamin D. Avasthi, Prachee The elusive actin cytoskeleton of a green alga expressing both conventional and divergent actins |
title | The elusive actin cytoskeleton of a green alga expressing both conventional and divergent actins |
title_full | The elusive actin cytoskeleton of a green alga expressing both conventional and divergent actins |
title_fullStr | The elusive actin cytoskeleton of a green alga expressing both conventional and divergent actins |
title_full_unstemmed | The elusive actin cytoskeleton of a green alga expressing both conventional and divergent actins |
title_short | The elusive actin cytoskeleton of a green alga expressing both conventional and divergent actins |
title_sort | elusive actin cytoskeleton of a green alga expressing both conventional and divergent actins |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6789165/ https://www.ncbi.nlm.nih.gov/pubmed/31532705 http://dx.doi.org/10.1091/mbc.E19-03-0141 |
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