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EGFR-RAS-MAPK signaling is confined to the plasma membrane and associated endorecycling protrusions

The subcellular localization of RAS GTPases defines the operational compartment of the EGFR-ERK1/2 signaling pathway within cells. Hence, we used live-cell imaging to demonstrate that endogenous KRAS and NRAS tagged with mNeonGreen are predominantly localized to the plasma membrane. NRAS was also pr...

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Detalles Bibliográficos
Autores principales: Surve, Sachin, Watkins, Simon C., Sorkin, Alexander
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Rockefeller University Press 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8563293/
https://www.ncbi.nlm.nih.gov/pubmed/34515735
http://dx.doi.org/10.1083/jcb.202107103
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author Surve, Sachin
Watkins, Simon C.
Sorkin, Alexander
author_facet Surve, Sachin
Watkins, Simon C.
Sorkin, Alexander
author_sort Surve, Sachin
collection PubMed
description The subcellular localization of RAS GTPases defines the operational compartment of the EGFR-ERK1/2 signaling pathway within cells. Hence, we used live-cell imaging to demonstrate that endogenous KRAS and NRAS tagged with mNeonGreen are predominantly localized to the plasma membrane. NRAS was also present in the Golgi apparatus and a tubular, plasma-membrane derived endorecycling compartment, enriched in recycling endosome markers (TERC). In EGF-stimulated cells, there was essentially no colocalization of either mNeonGreen-KRAS or mNeonGreen-NRAS with endosomal EGFR, which, by contrast, remained associated with endogenous Grb2-mNeonGreen, a receptor adaptor upstream of RAS. ERK1/2 activity was diminished by blocking cell surface EGFR with cetuximab, even after most ligand-bound, Grb2-associated EGFRs were internalized. Endogenous mCherry-tagged RAF1, an effector of RAS, was recruited to the plasma membrane, with subsequent accumulation in mNG-NRAS–containing TERCs. We propose that a small pool of surface EGFRs sustain signaling within the RAS-ERK1/2 pathway and that RAS activation persists in TERCs, whereas endosomal EGFR does not significantly contribute to ERK1/2 activity.
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spelling pubmed-85632932022-05-01 EGFR-RAS-MAPK signaling is confined to the plasma membrane and associated endorecycling protrusions Surve, Sachin Watkins, Simon C. Sorkin, Alexander J Cell Biol Article The subcellular localization of RAS GTPases defines the operational compartment of the EGFR-ERK1/2 signaling pathway within cells. Hence, we used live-cell imaging to demonstrate that endogenous KRAS and NRAS tagged with mNeonGreen are predominantly localized to the plasma membrane. NRAS was also present in the Golgi apparatus and a tubular, plasma-membrane derived endorecycling compartment, enriched in recycling endosome markers (TERC). In EGF-stimulated cells, there was essentially no colocalization of either mNeonGreen-KRAS or mNeonGreen-NRAS with endosomal EGFR, which, by contrast, remained associated with endogenous Grb2-mNeonGreen, a receptor adaptor upstream of RAS. ERK1/2 activity was diminished by blocking cell surface EGFR with cetuximab, even after most ligand-bound, Grb2-associated EGFRs were internalized. Endogenous mCherry-tagged RAF1, an effector of RAS, was recruited to the plasma membrane, with subsequent accumulation in mNG-NRAS–containing TERCs. We propose that a small pool of surface EGFRs sustain signaling within the RAS-ERK1/2 pathway and that RAS activation persists in TERCs, whereas endosomal EGFR does not significantly contribute to ERK1/2 activity. Rockefeller University Press 2021-09-13 /pmc/articles/PMC8563293/ /pubmed/34515735 http://dx.doi.org/10.1083/jcb.202107103 Text en © 2021 Surve et al. https://creativecommons.org/licenses/by-nc-sa/4.0/http://www.rupress.org/terms/This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/).
spellingShingle Article
Surve, Sachin
Watkins, Simon C.
Sorkin, Alexander
EGFR-RAS-MAPK signaling is confined to the plasma membrane and associated endorecycling protrusions
title EGFR-RAS-MAPK signaling is confined to the plasma membrane and associated endorecycling protrusions
title_full EGFR-RAS-MAPK signaling is confined to the plasma membrane and associated endorecycling protrusions
title_fullStr EGFR-RAS-MAPK signaling is confined to the plasma membrane and associated endorecycling protrusions
title_full_unstemmed EGFR-RAS-MAPK signaling is confined to the plasma membrane and associated endorecycling protrusions
title_short EGFR-RAS-MAPK signaling is confined to the plasma membrane and associated endorecycling protrusions
title_sort egfr-ras-mapk signaling is confined to the plasma membrane and associated endorecycling protrusions
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8563293/
https://www.ncbi.nlm.nih.gov/pubmed/34515735
http://dx.doi.org/10.1083/jcb.202107103
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