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Stress-dependent cell stiffening by tardigrade tolerance proteins that reversibly form a filamentous network and gel

Tardigrades are able to tolerate almost complete dehydration by entering a reversible ametabolic state called anhydrobiosis and resume their animation upon rehydration. Dehydrated tardigrades are exceptionally stable and withstand various physical extremes. Although trehalose and late embryogenesis...

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Autores principales: Tanaka, Akihiro, Nakano, Tomomi, Watanabe, Kento, Masuda, Kazutoshi, Honda, Gen, Kamata, Shuichi, Yasui, Reitaro, Kozuka-Hata, Hiroko, Watanabe, Chiho, Chinen, Takumi, Kitagawa, Daiju, Sawai, Satoshi, Oyama, Masaaki, Yanagisawa, Miho, Kunieda, Takekazu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9592077/
https://www.ncbi.nlm.nih.gov/pubmed/36067153
http://dx.doi.org/10.1371/journal.pbio.3001780
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author Tanaka, Akihiro
Nakano, Tomomi
Watanabe, Kento
Masuda, Kazutoshi
Honda, Gen
Kamata, Shuichi
Yasui, Reitaro
Kozuka-Hata, Hiroko
Watanabe, Chiho
Chinen, Takumi
Kitagawa, Daiju
Sawai, Satoshi
Oyama, Masaaki
Yanagisawa, Miho
Kunieda, Takekazu
author_facet Tanaka, Akihiro
Nakano, Tomomi
Watanabe, Kento
Masuda, Kazutoshi
Honda, Gen
Kamata, Shuichi
Yasui, Reitaro
Kozuka-Hata, Hiroko
Watanabe, Chiho
Chinen, Takumi
Kitagawa, Daiju
Sawai, Satoshi
Oyama, Masaaki
Yanagisawa, Miho
Kunieda, Takekazu
author_sort Tanaka, Akihiro
collection PubMed
description Tardigrades are able to tolerate almost complete dehydration by entering a reversible ametabolic state called anhydrobiosis and resume their animation upon rehydration. Dehydrated tardigrades are exceptionally stable and withstand various physical extremes. Although trehalose and late embryogenesis abundant (LEA) proteins have been extensively studied as potent protectants against dehydration in other anhydrobiotic organisms, tardigrades produce high amounts of tardigrade-unique protective proteins. Cytoplasmic-abundant heat-soluble (CAHS) proteins are uniquely invented in the lineage of eutardigrades, a major class of the phylum Tardigrada and are essential for their anhydrobiotic survival. However, the precise mechanisms of their action in this protective role are not fully understood. In the present study, we first postulated the presence of tolerance proteins that form protective condensates via phase separation in a stress-dependent manner and searched for tardigrade proteins that reversibly form condensates upon dehydration-like stress. Through a comprehensive search using a desolvating agent, trifluoroethanol (TFE), we identified 336 proteins, collectively dubbed “TFE-Dependent ReversiblY condensing Proteins (T-DRYPs).” Unexpectedly, we rediscovered CAHS proteins as highly enriched in T-DRYPs, 3 of which were major components of T-DRYPs. We revealed that these CAHS proteins reversibly polymerize into many cytoskeleton-like filaments depending on hyperosmotic stress in cultured cells and undergo reversible gel-transition in vitro. Furthermore, CAHS proteins increased cell stiffness in a hyperosmotic stress-dependent manner and counteract the cell shrinkage caused by osmotic pressure, and even improved the survival against hyperosmotic stress. The conserved putative helical C-terminal region is necessary and sufficient for filament formation by CAHS proteins, and mutations disrupting the secondary structure of this region impaired both the filament formation and the gel transition. On the basis of these results, we propose that CAHS proteins are novel cytoskeleton-like proteins that form filamentous networks and undergo gel-transition in a stress-dependent manner to provide on-demand physical stabilization of cell integrity against deformative forces during dehydration and could contribute to the exceptional physical stability in a dehydrated state.
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spelling pubmed-95920772022-10-25 Stress-dependent cell stiffening by tardigrade tolerance proteins that reversibly form a filamentous network and gel Tanaka, Akihiro Nakano, Tomomi Watanabe, Kento Masuda, Kazutoshi Honda, Gen Kamata, Shuichi Yasui, Reitaro Kozuka-Hata, Hiroko Watanabe, Chiho Chinen, Takumi Kitagawa, Daiju Sawai, Satoshi Oyama, Masaaki Yanagisawa, Miho Kunieda, Takekazu PLoS Biol Discovery Report Tardigrades are able to tolerate almost complete dehydration by entering a reversible ametabolic state called anhydrobiosis and resume their animation upon rehydration. Dehydrated tardigrades are exceptionally stable and withstand various physical extremes. Although trehalose and late embryogenesis abundant (LEA) proteins have been extensively studied as potent protectants against dehydration in other anhydrobiotic organisms, tardigrades produce high amounts of tardigrade-unique protective proteins. Cytoplasmic-abundant heat-soluble (CAHS) proteins are uniquely invented in the lineage of eutardigrades, a major class of the phylum Tardigrada and are essential for their anhydrobiotic survival. However, the precise mechanisms of their action in this protective role are not fully understood. In the present study, we first postulated the presence of tolerance proteins that form protective condensates via phase separation in a stress-dependent manner and searched for tardigrade proteins that reversibly form condensates upon dehydration-like stress. Through a comprehensive search using a desolvating agent, trifluoroethanol (TFE), we identified 336 proteins, collectively dubbed “TFE-Dependent ReversiblY condensing Proteins (T-DRYPs).” Unexpectedly, we rediscovered CAHS proteins as highly enriched in T-DRYPs, 3 of which were major components of T-DRYPs. We revealed that these CAHS proteins reversibly polymerize into many cytoskeleton-like filaments depending on hyperosmotic stress in cultured cells and undergo reversible gel-transition in vitro. Furthermore, CAHS proteins increased cell stiffness in a hyperosmotic stress-dependent manner and counteract the cell shrinkage caused by osmotic pressure, and even improved the survival against hyperosmotic stress. The conserved putative helical C-terminal region is necessary and sufficient for filament formation by CAHS proteins, and mutations disrupting the secondary structure of this region impaired both the filament formation and the gel transition. On the basis of these results, we propose that CAHS proteins are novel cytoskeleton-like proteins that form filamentous networks and undergo gel-transition in a stress-dependent manner to provide on-demand physical stabilization of cell integrity against deformative forces during dehydration and could contribute to the exceptional physical stability in a dehydrated state. Public Library of Science 2022-09-06 /pmc/articles/PMC9592077/ /pubmed/36067153 http://dx.doi.org/10.1371/journal.pbio.3001780 Text en © 2022 Tanaka et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://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 Discovery Report
Tanaka, Akihiro
Nakano, Tomomi
Watanabe, Kento
Masuda, Kazutoshi
Honda, Gen
Kamata, Shuichi
Yasui, Reitaro
Kozuka-Hata, Hiroko
Watanabe, Chiho
Chinen, Takumi
Kitagawa, Daiju
Sawai, Satoshi
Oyama, Masaaki
Yanagisawa, Miho
Kunieda, Takekazu
Stress-dependent cell stiffening by tardigrade tolerance proteins that reversibly form a filamentous network and gel
title Stress-dependent cell stiffening by tardigrade tolerance proteins that reversibly form a filamentous network and gel
title_full Stress-dependent cell stiffening by tardigrade tolerance proteins that reversibly form a filamentous network and gel
title_fullStr Stress-dependent cell stiffening by tardigrade tolerance proteins that reversibly form a filamentous network and gel
title_full_unstemmed Stress-dependent cell stiffening by tardigrade tolerance proteins that reversibly form a filamentous network and gel
title_short Stress-dependent cell stiffening by tardigrade tolerance proteins that reversibly form a filamentous network and gel
title_sort stress-dependent cell stiffening by tardigrade tolerance proteins that reversibly form a filamentous network and gel
topic Discovery Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9592077/
https://www.ncbi.nlm.nih.gov/pubmed/36067153
http://dx.doi.org/10.1371/journal.pbio.3001780
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