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Pil1 cytoplasmic rods contain bundles of crosslinked tubules

Cytoskeletal polymers are organized into a wide variety of higher-order structures in cells. The yeast BAR domain protein Pil1 self-assembles into tubules in vitro, and forms linear polymers at cortical eisosomes in cells. In the fission yeast S. pombe, over-expressed Pil1 forms thick rods that deta...

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Autores principales: Kabeche, Ruth, Howard, Louisa, Moseley, James B
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Taylor & Francis 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4594478/
https://www.ncbi.nlm.nih.gov/pubmed/26609339
http://dx.doi.org/10.4161/19420889.2014.990848
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author Kabeche, Ruth
Howard, Louisa
Moseley, James B
author_facet Kabeche, Ruth
Howard, Louisa
Moseley, James B
author_sort Kabeche, Ruth
collection PubMed
description Cytoskeletal polymers are organized into a wide variety of higher-order structures in cells. The yeast BAR domain protein Pil1 self-assembles into tubules in vitro, and forms linear polymers at cortical eisosomes in cells. In the fission yeast S. pombe, over-expressed Pil1 forms thick rods that detach from the plasma membrane. In this study, we used thin-section electron microscopy to determine the ultrastructure of these cytoplasmic Pil1 rods. We found that cytoplasmic rods contained crosslinked Pil1 tubules that displayed regular, hexagonal spacing. These bundles were stained by filipin, a sterol-binding fluorescent dye, suggesting that they contained lipids. Cytoplasmic Pil1 rods were present but less abundant in sle1Δ and fhn1Δ mutant cells. We also found that endogenous Pil1 formed thick rods under saturated growth conditions. Taken together, our findings suggest the presence of cellular mechanisms that assemble Pil1 tubules into higher-order structures.
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spelling pubmed-45944782015-11-25 Pil1 cytoplasmic rods contain bundles of crosslinked tubules Kabeche, Ruth Howard, Louisa Moseley, James B Commun Integr Biol Short Communication Cytoskeletal polymers are organized into a wide variety of higher-order structures in cells. The yeast BAR domain protein Pil1 self-assembles into tubules in vitro, and forms linear polymers at cortical eisosomes in cells. In the fission yeast S. pombe, over-expressed Pil1 forms thick rods that detach from the plasma membrane. In this study, we used thin-section electron microscopy to determine the ultrastructure of these cytoplasmic Pil1 rods. We found that cytoplasmic rods contained crosslinked Pil1 tubules that displayed regular, hexagonal spacing. These bundles were stained by filipin, a sterol-binding fluorescent dye, suggesting that they contained lipids. Cytoplasmic Pil1 rods were present but less abundant in sle1Δ and fhn1Δ mutant cells. We also found that endogenous Pil1 formed thick rods under saturated growth conditions. Taken together, our findings suggest the presence of cellular mechanisms that assemble Pil1 tubules into higher-order structures. Taylor & Francis 2015-03-04 /pmc/articles/PMC4594478/ /pubmed/26609339 http://dx.doi.org/10.4161/19420889.2014.990848 Text en © 2015 The Author(s). Published with license by Taylor & Francis Group, LLC http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Short Communication
Kabeche, Ruth
Howard, Louisa
Moseley, James B
Pil1 cytoplasmic rods contain bundles of crosslinked tubules
title Pil1 cytoplasmic rods contain bundles of crosslinked tubules
title_full Pil1 cytoplasmic rods contain bundles of crosslinked tubules
title_fullStr Pil1 cytoplasmic rods contain bundles of crosslinked tubules
title_full_unstemmed Pil1 cytoplasmic rods contain bundles of crosslinked tubules
title_short Pil1 cytoplasmic rods contain bundles of crosslinked tubules
title_sort pil1 cytoplasmic rods contain bundles of crosslinked tubules
topic Short Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4594478/
https://www.ncbi.nlm.nih.gov/pubmed/26609339
http://dx.doi.org/10.4161/19420889.2014.990848
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