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Membrane stretching activates calcium permeability of a putative channel Pkd2 during fission yeast cytokinesis

Pkd2 is the fission yeast homologue of polycystins. This putative ion channel localizes to the plasma membrane. It is required for the expansion of cell volume during interphase growth and cytokinesis, the last step of cell division. However, the channel activity of Pkd2 remains untested. Here, we e...

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Autores principales: Poddar, Abhishek, Hsu, Yen-Yu, Zhang, Faith, Shamma, Abeda, Kreais, Zachary, Muller, Clare, Malla, Mamata, Ray, Aniruddha, Liu, Allen P., Chen, Qian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9727806/
https://www.ncbi.nlm.nih.gov/pubmed/36200871
http://dx.doi.org/10.1091/mbc.E22-07-0248
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author Poddar, Abhishek
Hsu, Yen-Yu
Zhang, Faith
Shamma, Abeda
Kreais, Zachary
Muller, Clare
Malla, Mamata
Ray, Aniruddha
Liu, Allen P.
Chen, Qian
author_facet Poddar, Abhishek
Hsu, Yen-Yu
Zhang, Faith
Shamma, Abeda
Kreais, Zachary
Muller, Clare
Malla, Mamata
Ray, Aniruddha
Liu, Allen P.
Chen, Qian
author_sort Poddar, Abhishek
collection PubMed
description Pkd2 is the fission yeast homologue of polycystins. This putative ion channel localizes to the plasma membrane. It is required for the expansion of cell volume during interphase growth and cytokinesis, the last step of cell division. However, the channel activity of Pkd2 remains untested. Here, we examined the calcium permeability and mechanosensitivity of Pkd2 through in vitro reconstitution and calcium imaging of pkd2 mutant cells. Pkd2 was translated and inserted into the lipid bilayers of giant unilamellar vesicles using a cell-free expression system. The reconstituted Pkd2 permeated calcium when the membrane was stretched via hypoosmotic shock. In vivo, inactivation of Pkd2 through a temperature-sensitive mutation pkd2-B42 reduced the average intracellular calcium level by 34%. Compared with the wild type, the hypomorphic mutation pkd2-81KD reduced the amplitude of hypoosmotic shock–triggered calcium spikes by 59%. During cytokinesis, mutations of pkd2 reduced the calcium spikes, accompanying cell separation and the ensuing membrane stretching, by 60%. We concluded that fission yeast polycystin Pkd2 allows calcium influx when activated by membrane stretching, representing a likely mechanosensitive channel that contributes to the cytokinetic calcium spikes.
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spelling pubmed-97278062023-02-02 Membrane stretching activates calcium permeability of a putative channel Pkd2 during fission yeast cytokinesis Poddar, Abhishek Hsu, Yen-Yu Zhang, Faith Shamma, Abeda Kreais, Zachary Muller, Clare Malla, Mamata Ray, Aniruddha Liu, Allen P. Chen, Qian Mol Biol Cell Articles Pkd2 is the fission yeast homologue of polycystins. This putative ion channel localizes to the plasma membrane. It is required for the expansion of cell volume during interphase growth and cytokinesis, the last step of cell division. However, the channel activity of Pkd2 remains untested. Here, we examined the calcium permeability and mechanosensitivity of Pkd2 through in vitro reconstitution and calcium imaging of pkd2 mutant cells. Pkd2 was translated and inserted into the lipid bilayers of giant unilamellar vesicles using a cell-free expression system. The reconstituted Pkd2 permeated calcium when the membrane was stretched via hypoosmotic shock. In vivo, inactivation of Pkd2 through a temperature-sensitive mutation pkd2-B42 reduced the average intracellular calcium level by 34%. Compared with the wild type, the hypomorphic mutation pkd2-81KD reduced the amplitude of hypoosmotic shock–triggered calcium spikes by 59%. During cytokinesis, mutations of pkd2 reduced the calcium spikes, accompanying cell separation and the ensuing membrane stretching, by 60%. We concluded that fission yeast polycystin Pkd2 allows calcium influx when activated by membrane stretching, representing a likely mechanosensitive channel that contributes to the cytokinetic calcium spikes. The American Society for Cell Biology 2022-11-18 /pmc/articles/PMC9727806/ /pubmed/36200871 http://dx.doi.org/10.1091/mbc.E22-07-0248 Text en © 2022 Poddar 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. https://creativecommons.org/licenses/by-nc-sa/4.0/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 4.0 International Creative Commons License.
spellingShingle Articles
Poddar, Abhishek
Hsu, Yen-Yu
Zhang, Faith
Shamma, Abeda
Kreais, Zachary
Muller, Clare
Malla, Mamata
Ray, Aniruddha
Liu, Allen P.
Chen, Qian
Membrane stretching activates calcium permeability of a putative channel Pkd2 during fission yeast cytokinesis
title Membrane stretching activates calcium permeability of a putative channel Pkd2 during fission yeast cytokinesis
title_full Membrane stretching activates calcium permeability of a putative channel Pkd2 during fission yeast cytokinesis
title_fullStr Membrane stretching activates calcium permeability of a putative channel Pkd2 during fission yeast cytokinesis
title_full_unstemmed Membrane stretching activates calcium permeability of a putative channel Pkd2 during fission yeast cytokinesis
title_short Membrane stretching activates calcium permeability of a putative channel Pkd2 during fission yeast cytokinesis
title_sort membrane stretching activates calcium permeability of a putative channel pkd2 during fission yeast cytokinesis
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9727806/
https://www.ncbi.nlm.nih.gov/pubmed/36200871
http://dx.doi.org/10.1091/mbc.E22-07-0248
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