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High-throughput, single-particle tracking reveals nested membrane domains that dictate KRas(G12D) diffusion and trafficking

Membrane nanodomains have been implicated in Ras signaling, but what these domains are and how they interact with Ras remain obscure. Here, using single particle tracking with photoactivated localization microscopy (spt-PALM) and detailed trajectory analysis, we show that distinct membrane domains d...

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Autores principales: Lee, Yerim, Phelps, Carey, Huang, Tao, Mostofian, Barmak, Wu, Lei, Zhang, Ying, Tao, Kai, Chang, Young Hwan, Stork, Philip JS, Gray, Joe W, Zuckerman, Daniel M, Nan, Xiaolin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7060040/
https://www.ncbi.nlm.nih.gov/pubmed/31674905
http://dx.doi.org/10.7554/eLife.46393
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author Lee, Yerim
Phelps, Carey
Huang, Tao
Mostofian, Barmak
Wu, Lei
Zhang, Ying
Tao, Kai
Chang, Young Hwan
Stork, Philip JS
Gray, Joe W
Zuckerman, Daniel M
Nan, Xiaolin
author_facet Lee, Yerim
Phelps, Carey
Huang, Tao
Mostofian, Barmak
Wu, Lei
Zhang, Ying
Tao, Kai
Chang, Young Hwan
Stork, Philip JS
Gray, Joe W
Zuckerman, Daniel M
Nan, Xiaolin
author_sort Lee, Yerim
collection PubMed
description Membrane nanodomains have been implicated in Ras signaling, but what these domains are and how they interact with Ras remain obscure. Here, using single particle tracking with photoactivated localization microscopy (spt-PALM) and detailed trajectory analysis, we show that distinct membrane domains dictate KRas(G12D) (an active KRas mutant) diffusion and trafficking in U2OS cells. KRas(G12D) exhibits an immobile state in ~70 nm domains, each embedded in a larger domain (~200 nm) that confers intermediate mobility, while the rest of the membrane supports fast diffusion. Moreover, KRas(G12D) is continuously removed from the membrane via the immobile state and replenished to the fast state, reminiscent of Ras internalization and recycling. Importantly, both the diffusion and trafficking properties of KRas(G12D) remain invariant over a broad range of protein expression levels. Our results reveal how membrane organization dictates membrane diffusion and trafficking of Ras and offer new insight into the spatial regulation of Ras signaling.
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spelling pubmed-70600402020-03-09 High-throughput, single-particle tracking reveals nested membrane domains that dictate KRas(G12D) diffusion and trafficking Lee, Yerim Phelps, Carey Huang, Tao Mostofian, Barmak Wu, Lei Zhang, Ying Tao, Kai Chang, Young Hwan Stork, Philip JS Gray, Joe W Zuckerman, Daniel M Nan, Xiaolin eLife Physics of Living Systems Membrane nanodomains have been implicated in Ras signaling, but what these domains are and how they interact with Ras remain obscure. Here, using single particle tracking with photoactivated localization microscopy (spt-PALM) and detailed trajectory analysis, we show that distinct membrane domains dictate KRas(G12D) (an active KRas mutant) diffusion and trafficking in U2OS cells. KRas(G12D) exhibits an immobile state in ~70 nm domains, each embedded in a larger domain (~200 nm) that confers intermediate mobility, while the rest of the membrane supports fast diffusion. Moreover, KRas(G12D) is continuously removed from the membrane via the immobile state and replenished to the fast state, reminiscent of Ras internalization and recycling. Importantly, both the diffusion and trafficking properties of KRas(G12D) remain invariant over a broad range of protein expression levels. Our results reveal how membrane organization dictates membrane diffusion and trafficking of Ras and offer new insight into the spatial regulation of Ras signaling. eLife Sciences Publications, Ltd 2019-11-01 /pmc/articles/PMC7060040/ /pubmed/31674905 http://dx.doi.org/10.7554/eLife.46393 Text en © 2019, Lee et al http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Physics of Living Systems
Lee, Yerim
Phelps, Carey
Huang, Tao
Mostofian, Barmak
Wu, Lei
Zhang, Ying
Tao, Kai
Chang, Young Hwan
Stork, Philip JS
Gray, Joe W
Zuckerman, Daniel M
Nan, Xiaolin
High-throughput, single-particle tracking reveals nested membrane domains that dictate KRas(G12D) diffusion and trafficking
title High-throughput, single-particle tracking reveals nested membrane domains that dictate KRas(G12D) diffusion and trafficking
title_full High-throughput, single-particle tracking reveals nested membrane domains that dictate KRas(G12D) diffusion and trafficking
title_fullStr High-throughput, single-particle tracking reveals nested membrane domains that dictate KRas(G12D) diffusion and trafficking
title_full_unstemmed High-throughput, single-particle tracking reveals nested membrane domains that dictate KRas(G12D) diffusion and trafficking
title_short High-throughput, single-particle tracking reveals nested membrane domains that dictate KRas(G12D) diffusion and trafficking
title_sort high-throughput, single-particle tracking reveals nested membrane domains that dictate kras(g12d) diffusion and trafficking
topic Physics of Living Systems
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7060040/
https://www.ncbi.nlm.nih.gov/pubmed/31674905
http://dx.doi.org/10.7554/eLife.46393
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