An optogenetic system to control membrane phospholipid asymmetry through flippase activation in budding yeast

Abstract Lipid asymmetry in biological membranes is essential for various cell functions, such as cell polarity, cytokinesis, and apoptosis. P4-ATPases (flippases) are involved in the generation of such asymmetry. In Saccharomyces cerevisiae, the protein kinases Fpk1p/Fpk2p activate the P4-ATPases D...

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Autores principales:	Tomomi Suzuki, Tetsuo Mioka, Kazuma Tanaka, Akira Nagatani
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/74a8bd5f78ee4df692717e1bdefea086
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spelling	oai:doaj.org-article:74a8bd5f78ee4df692717e1bdefea0862021-12-02T16:06:39ZAn optogenetic system to control membrane phospholipid asymmetry through flippase activation in budding yeast10.1038/s41598-020-69459-02045-2322https://doaj.org/article/74a8bd5f78ee4df692717e1bdefea0862020-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-69459-0https://doaj.org/toc/2045-2322Abstract Lipid asymmetry in biological membranes is essential for various cell functions, such as cell polarity, cytokinesis, and apoptosis. P4-ATPases (flippases) are involved in the generation of such asymmetry. In Saccharomyces cerevisiae, the protein kinases Fpk1p/Fpk2p activate the P4-ATPases Dnf1p/Dnf2p by phosphorylation. Previously, we have shown that a blue-light-dependent protein kinase, phototropin from Chlamydomonas reinhardtii (CrPHOT), complements defects in an fpk1Δ fpk2Δ mutant. Herein, we investigated whether CrPHOT optically regulates P4-ATPase activity. First, we demonstrated that the translocation of NBD-labelled phospholipids to the cytoplasmic leaflet via P4-ATPases was promoted by blue-light irradiation in fpk1Δ fpk2Δ cells with CrPHOT. In addition, blue light completely suppressed the defects in membrane functions (such as endocytic recycling, actin depolarization, and apical-isotropic growth switching) caused by fpk1Δ fpk2Δ mutations. All responses required the kinase activity of CrPHOT. Hence, these results indicate the utility of CrPHOT as a powerful and first tool for optogenetic manipulation of P4-ATPase activity.Tomomi SuzukiTetsuo MiokaKazuma TanakaAkira NagataniNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-14 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Tomomi Suzuki Tetsuo Mioka Kazuma Tanaka Akira Nagatani An optogenetic system to control membrane phospholipid asymmetry through flippase activation in budding yeast
description	Abstract Lipid asymmetry in biological membranes is essential for various cell functions, such as cell polarity, cytokinesis, and apoptosis. P4-ATPases (flippases) are involved in the generation of such asymmetry. In Saccharomyces cerevisiae, the protein kinases Fpk1p/Fpk2p activate the P4-ATPases Dnf1p/Dnf2p by phosphorylation. Previously, we have shown that a blue-light-dependent protein kinase, phototropin from Chlamydomonas reinhardtii (CrPHOT), complements defects in an fpk1Δ fpk2Δ mutant. Herein, we investigated whether CrPHOT optically regulates P4-ATPase activity. First, we demonstrated that the translocation of NBD-labelled phospholipids to the cytoplasmic leaflet via P4-ATPases was promoted by blue-light irradiation in fpk1Δ fpk2Δ cells with CrPHOT. In addition, blue light completely suppressed the defects in membrane functions (such as endocytic recycling, actin depolarization, and apical-isotropic growth switching) caused by fpk1Δ fpk2Δ mutations. All responses required the kinase activity of CrPHOT. Hence, these results indicate the utility of CrPHOT as a powerful and first tool for optogenetic manipulation of P4-ATPase activity.
format	article
author	Tomomi Suzuki Tetsuo Mioka Kazuma Tanaka Akira Nagatani
author_facet	Tomomi Suzuki Tetsuo Mioka Kazuma Tanaka Akira Nagatani
author_sort	Tomomi Suzuki
title	An optogenetic system to control membrane phospholipid asymmetry through flippase activation in budding yeast
title_short	An optogenetic system to control membrane phospholipid asymmetry through flippase activation in budding yeast
title_full	An optogenetic system to control membrane phospholipid asymmetry through flippase activation in budding yeast
title_fullStr	An optogenetic system to control membrane phospholipid asymmetry through flippase activation in budding yeast
title_full_unstemmed	An optogenetic system to control membrane phospholipid asymmetry through flippase activation in budding yeast
title_sort	optogenetic system to control membrane phospholipid asymmetry through flippase activation in budding yeast
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/74a8bd5f78ee4df692717e1bdefea086
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An optogenetic system to control membrane phospholipid asymmetry through flippase activation in budding yeast

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