PKCα diffusion and translocation are independent of an intact cytoskeleton

Abstract Translocation of cytosolic cPKC to the plasma membrane is a key event in their activation process but its exact nature is still unclear with particular dispute whether sole diffusion or additional active transport along the cell’s cytoskeleton contributes to cPKC’s dynamics. This was addres...

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Autores principales:	Xin Hui, Benjamin Sauer, Lars Kaestner, Karsten Kruse, Peter Lipp
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/8856c3bf78d947cc81b72d7b74e956a0
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spelling	oai:doaj.org-article:8856c3bf78d947cc81b72d7b74e956a02021-12-02T12:32:50ZPKCα diffusion and translocation are independent of an intact cytoskeleton10.1038/s41598-017-00560-72045-2322https://doaj.org/article/8856c3bf78d947cc81b72d7b74e956a02017-03-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-00560-7https://doaj.org/toc/2045-2322Abstract Translocation of cytosolic cPKC to the plasma membrane is a key event in their activation process but its exact nature is still unclear with particular dispute whether sole diffusion or additional active transport along the cell’s cytoskeleton contributes to cPKC’s dynamics. This was addressed by analyzing the recruitment behavior of PKCα while manipulating the cytoskeleton. Photolytic Ca2+ uncaging allowed us to quantify the kinetics of PKCα redistribution to the plasma membrane when fused to monomeric, dimeric and tetrameric fluorescence proteins. Results indicated that translocation kinetics were modulated by the state of oligomerization as expected for varying Stokes’ radii of the participating proteins. Following depolymerization of the microtubules and the actin filaments we found that Ca2+ induced membrane accumulation of PKCα was independent of the filamentous state of the cytoskeleton. Fusion of PKCα to the photo-convertible fluorescent protein Dendra2 enabled the investigation of PKCα-cytoskeleton interactions under resting conditions. Redistribution following spatially restricted photoconversion showed that the mobility of the fusion protein was independent of the state of the cytoskeleton. Our data demonstrated that in living cells neither actin filaments nor microtubules contribute to PKCα’s cytosolic mobility or Ca2+-induced translocation to the plasma membrane. Instead translocation is a solely diffusion-driven process.Xin HuiBenjamin SauerLars KaestnerKarsten KrusePeter LippNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Xin Hui Benjamin Sauer Lars Kaestner Karsten Kruse Peter Lipp PKCα diffusion and translocation are independent of an intact cytoskeleton
description	Abstract Translocation of cytosolic cPKC to the plasma membrane is a key event in their activation process but its exact nature is still unclear with particular dispute whether sole diffusion or additional active transport along the cell’s cytoskeleton contributes to cPKC’s dynamics. This was addressed by analyzing the recruitment behavior of PKCα while manipulating the cytoskeleton. Photolytic Ca2+ uncaging allowed us to quantify the kinetics of PKCα redistribution to the plasma membrane when fused to monomeric, dimeric and tetrameric fluorescence proteins. Results indicated that translocation kinetics were modulated by the state of oligomerization as expected for varying Stokes’ radii of the participating proteins. Following depolymerization of the microtubules and the actin filaments we found that Ca2+ induced membrane accumulation of PKCα was independent of the filamentous state of the cytoskeleton. Fusion of PKCα to the photo-convertible fluorescent protein Dendra2 enabled the investigation of PKCα-cytoskeleton interactions under resting conditions. Redistribution following spatially restricted photoconversion showed that the mobility of the fusion protein was independent of the state of the cytoskeleton. Our data demonstrated that in living cells neither actin filaments nor microtubules contribute to PKCα’s cytosolic mobility or Ca2+-induced translocation to the plasma membrane. Instead translocation is a solely diffusion-driven process.
format	article
author	Xin Hui Benjamin Sauer Lars Kaestner Karsten Kruse Peter Lipp
author_facet	Xin Hui Benjamin Sauer Lars Kaestner Karsten Kruse Peter Lipp
author_sort	Xin Hui
title	PKCα diffusion and translocation are independent of an intact cytoskeleton
title_short	PKCα diffusion and translocation are independent of an intact cytoskeleton
title_full	PKCα diffusion and translocation are independent of an intact cytoskeleton
title_fullStr	PKCα diffusion and translocation are independent of an intact cytoskeleton
title_full_unstemmed	PKCα diffusion and translocation are independent of an intact cytoskeleton
title_sort	pkcα diffusion and translocation are independent of an intact cytoskeleton
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/8856c3bf78d947cc81b72d7b74e956a0
work_keys_str_mv	AT xinhui pkcadiffusionandtranslocationareindependentofanintactcytoskeleton AT benjaminsauer pkcadiffusionandtranslocationareindependentofanintactcytoskeleton AT larskaestner pkcadiffusionandtranslocationareindependentofanintactcytoskeleton AT karstenkruse pkcadiffusionandtranslocationareindependentofanintactcytoskeleton AT peterlipp pkcadiffusionandtranslocationareindependentofanintactcytoskeleton
_version_	1718393930890870784

PKCα diffusion and translocation are independent of an intact cytoskeleton

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