Quantitative characterization of the auxin-inducible degron: a guide for dynamic protein depletion in single yeast cells

Abstract Perturbations are essential for the interrogation of biological systems. The auxin-inducible degron harbors great potential for dynamic protein depletion in yeast. Here, we thoroughly and quantitatively characterize the auxin-inducible degron in single yeast cells. We show that an auxin con...

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Autores principales:	Alexandros Papagiannakis, Janeska J de Jonge, Zheng Zhang, Matthias Heinemann
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/4d1fe0daf87c4d53ae8b2958de3386dc
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spelling	oai:doaj.org-article:4d1fe0daf87c4d53ae8b2958de3386dc2021-12-02T11:52:27ZQuantitative characterization of the auxin-inducible degron: a guide for dynamic protein depletion in single yeast cells10.1038/s41598-017-04791-62045-2322https://doaj.org/article/4d1fe0daf87c4d53ae8b2958de3386dc2017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-04791-6https://doaj.org/toc/2045-2322Abstract Perturbations are essential for the interrogation of biological systems. The auxin-inducible degron harbors great potential for dynamic protein depletion in yeast. Here, we thoroughly and quantitatively characterize the auxin-inducible degron in single yeast cells. We show that an auxin concentration of 0.25 mM is necessary for fast and uniform protein depletion between single cells, and that in mother cells proteins are depleted faster than their daughters. Although, protein recovery starts immediately after removal of auxin, it takes multiple generations before equilibrium is reached between protein synthesis and dilution, which is when the original protein levels are restored. Further, we found that blue light, used for GFP excitation, together with auxin results in growth defects, caused by the photo-destruction of auxin to its toxic derivatives, which can be avoided if indole-free auxin substitutes are used. Our work provides guidelines for the successful combination of microscopy, microfluidics and the auxin-inducible degron, offering the yeast community an unprecedented tool for dynamic perturbations on the single cell level.Alexandros PapagiannakisJaneska J de JongeZheng ZhangMatthias HeinemannNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-13 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Alexandros Papagiannakis Janeska J de Jonge Zheng Zhang Matthias Heinemann Quantitative characterization of the auxin-inducible degron: a guide for dynamic protein depletion in single yeast cells
description	Abstract Perturbations are essential for the interrogation of biological systems. The auxin-inducible degron harbors great potential for dynamic protein depletion in yeast. Here, we thoroughly and quantitatively characterize the auxin-inducible degron in single yeast cells. We show that an auxin concentration of 0.25 mM is necessary for fast and uniform protein depletion between single cells, and that in mother cells proteins are depleted faster than their daughters. Although, protein recovery starts immediately after removal of auxin, it takes multiple generations before equilibrium is reached between protein synthesis and dilution, which is when the original protein levels are restored. Further, we found that blue light, used for GFP excitation, together with auxin results in growth defects, caused by the photo-destruction of auxin to its toxic derivatives, which can be avoided if indole-free auxin substitutes are used. Our work provides guidelines for the successful combination of microscopy, microfluidics and the auxin-inducible degron, offering the yeast community an unprecedented tool for dynamic perturbations on the single cell level.
format	article
author	Alexandros Papagiannakis Janeska J de Jonge Zheng Zhang Matthias Heinemann
author_facet	Alexandros Papagiannakis Janeska J de Jonge Zheng Zhang Matthias Heinemann
author_sort	Alexandros Papagiannakis
title	Quantitative characterization of the auxin-inducible degron: a guide for dynamic protein depletion in single yeast cells
title_short	Quantitative characterization of the auxin-inducible degron: a guide for dynamic protein depletion in single yeast cells
title_full	Quantitative characterization of the auxin-inducible degron: a guide for dynamic protein depletion in single yeast cells
title_fullStr	Quantitative characterization of the auxin-inducible degron: a guide for dynamic protein depletion in single yeast cells
title_full_unstemmed	Quantitative characterization of the auxin-inducible degron: a guide for dynamic protein depletion in single yeast cells
title_sort	quantitative characterization of the auxin-inducible degron: a guide for dynamic protein depletion in single yeast cells
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/4d1fe0daf87c4d53ae8b2958de3386dc
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Quantitative characterization of the auxin-inducible degron: a guide for dynamic protein depletion in single yeast cells

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