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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...

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Autores principales: Craig, Evan W., Mueller, David M., Bigge, Brae M., Schaffer, Miroslava, Engel, Benjamin D., Avasthi, Prachee
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2019
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.
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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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