@article{beec78b0f0a74b9eb7fcef55b29e1379,
title = "High-throughput single-particle tracking reveals 1 nested membrane domains that dictate krasg12d 2 diffusion and trafficking",
abstract = "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 KRasG12D (an active KRas mutant) diffusion and trafficking in U2OS cells. KRasG12D 23 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, KRasG12D 25 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 KRasG12D 28 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.",
author = "Yerim Lee and Carey Phelps and Tao Huang and Barmak Mostofian and Lei Wu and Ying Zhang and Kai Tao and Chang, {Young Hwan} and Stork, {Philip J.S.} and Gray, {Joe W.} and Zuckerman, {Daniel M.} and Xiaolin Nan",
note = "Funding Information: Consortium (CSBC) grant (U54 CA209988, PI: Joe Gray) and by the Knight Caner Early Detection Funding Information: Systems Biomedicine (OCSSB). DMZ acknowledges support from the OCSSB and the National Science Funding Information: The authors would like to thank many colleagues for their helpful discussions, including those at OHSU (Drs. Laura Heiser, Xubo Song, Molly Kulesz-Martin, Pamela Cassidy, and others) and at the Frederick National Laboratory for Cancer Research (Drs. Frank McCormick (also at UCSF), Thomas Turbyville, Dwight Nissley, and others). Research in the Nan lab was supported by startup funds from the Knight Cancer Institute at OHSU, the Damon Runyon Cancer Research Foundation, the M. J. Murdock Charitable Trust, and the Prospect Creek Foundation. XN is also supported by a Cancer Systems Biology Consortium (CSBC) grant (U54 CA209988, PI: Joe Gray) and by the Knight Caner Early Detection Advanced Research (CEDAR) Center at OHSU. YL is currently supported by OHSU Center for Spatial Systems Biomedicine (OCSSB). DMZ acknowledges support from the OCSSB and the National Science Foundation under grant MCB 1715823. We also thank Ms. Julia Shangguan (Nan Lab) for graphic illustrations and Dr. Martin Lind?n (University of Uppsala) for helpful discussions on vbSPT. Funding Information: Turbyville, Dwight Nissley, and others). Research in the Nan lab was supported by startup funds from the Funding Information: Charitable Trust, and the Prospect Creek Foundation. XN is also supported by a Cancer Systems Biology Funding Information: Advanced Research (CEDAR) Center at OHSU. YL is currently supported by OHSU Center for Spatial Publisher Copyright: {\textcopyright} 2019, eLife Sciences Publications Ltd. All rights reserved.",
year = "2019",
month = nov,
doi = "10.7554/eLife.46393",
language = "English (US)",
volume = "8",
journal = "eLife",
issn = "2050-084X",
publisher = "eLife Sciences Publications",
}