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Epidermal Growth Factor and Membrane Trafficking: Egf Receptor Activation of Endocytosis Requires Rab5a

Activated epidermal growth factor receptors recruit various intracellular proteins leading to signal generation and endocytic trafficking. Although activated receptors are rapidly internalized into the endocytic compartment and subsequently degraded in lysosomes, the linkage between signaling and en...

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Autores principales: Barbieri, M. Alejandro, Roberts, Richard L., Gumusboga, Aysel, Highfield, Hilary, Alvarez-Dominguez, Carmen, Wells, Alan, Stahl, Philip D.
Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 2000
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2185585/
https://www.ncbi.nlm.nih.gov/pubmed/11062256
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author Barbieri, M. Alejandro
Roberts, Richard L.
Gumusboga, Aysel
Highfield, Hilary
Alvarez-Dominguez, Carmen
Wells, Alan
Stahl, Philip D.
author_facet Barbieri, M. Alejandro
Roberts, Richard L.
Gumusboga, Aysel
Highfield, Hilary
Alvarez-Dominguez, Carmen
Wells, Alan
Stahl, Philip D.
author_sort Barbieri, M. Alejandro
collection PubMed
description Activated epidermal growth factor receptors recruit various intracellular proteins leading to signal generation and endocytic trafficking. Although activated receptors are rapidly internalized into the endocytic compartment and subsequently degraded in lysosomes, the linkage between signaling and endocytosis is not well understood. Here we show that EGF stimulation of NR6 cells induces a specific, rapid and transient activation of Rab5a. EGF also enhanced translocation of the Rab5 effector, early endosomal autoantigen 1 (EEA1), from cytosol to membrane. The activation of endocytosis, fluid phase and receptor mediated, by EGF was enhanced by Rab5a expression, but not by Rab5b, Rab5c, or Rab5a truncated at the NH(2) and/or COOH terminus. Dominant negative Rab5a (Rab5:N34) blocked EGF-stimulated receptor-mediated and fluid-phase endocytosis. EGF activation of Rab5a function was dependent on tyrosine residues in the COOH-terminal domain of the EGF receptor (EGFR). Removal of the entire COOH terminus by truncation (c'973 and c'991) abrogated ligand-induced Rab5a activation of endocytosis. A “kinase-dead” EGFR failed to stimulate Rab5a function. However, another EGF receptor mutant (c'1000), with the kinase domain intact and a single autophosphorylation site effectively signaled Rab5 activation. These results indicate that EGFR and Rab5a are linked via a cascade that results in the activation of Rab5a and that appears essential for internalization. The results point to an interdependent relationship between receptor activation, signal generation and endocytosis.
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spelling pubmed-21855852008-05-01 Epidermal Growth Factor and Membrane Trafficking: Egf Receptor Activation of Endocytosis Requires Rab5a Barbieri, M. Alejandro Roberts, Richard L. Gumusboga, Aysel Highfield, Hilary Alvarez-Dominguez, Carmen Wells, Alan Stahl, Philip D. J Cell Biol Original Article Activated epidermal growth factor receptors recruit various intracellular proteins leading to signal generation and endocytic trafficking. Although activated receptors are rapidly internalized into the endocytic compartment and subsequently degraded in lysosomes, the linkage between signaling and endocytosis is not well understood. Here we show that EGF stimulation of NR6 cells induces a specific, rapid and transient activation of Rab5a. EGF also enhanced translocation of the Rab5 effector, early endosomal autoantigen 1 (EEA1), from cytosol to membrane. The activation of endocytosis, fluid phase and receptor mediated, by EGF was enhanced by Rab5a expression, but not by Rab5b, Rab5c, or Rab5a truncated at the NH(2) and/or COOH terminus. Dominant negative Rab5a (Rab5:N34) blocked EGF-stimulated receptor-mediated and fluid-phase endocytosis. EGF activation of Rab5a function was dependent on tyrosine residues in the COOH-terminal domain of the EGF receptor (EGFR). Removal of the entire COOH terminus by truncation (c'973 and c'991) abrogated ligand-induced Rab5a activation of endocytosis. A “kinase-dead” EGFR failed to stimulate Rab5a function. However, another EGF receptor mutant (c'1000), with the kinase domain intact and a single autophosphorylation site effectively signaled Rab5 activation. These results indicate that EGFR and Rab5a are linked via a cascade that results in the activation of Rab5a and that appears essential for internalization. The results point to an interdependent relationship between receptor activation, signal generation and endocytosis. The Rockefeller University Press 2000-10-30 /pmc/articles/PMC2185585/ /pubmed/11062256 Text en © 2000 The Rockefeller University Press 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 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).
spellingShingle Original Article
Barbieri, M. Alejandro
Roberts, Richard L.
Gumusboga, Aysel
Highfield, Hilary
Alvarez-Dominguez, Carmen
Wells, Alan
Stahl, Philip D.
Epidermal Growth Factor and Membrane Trafficking: Egf Receptor Activation of Endocytosis Requires Rab5a
title Epidermal Growth Factor and Membrane Trafficking: Egf Receptor Activation of Endocytosis Requires Rab5a
title_full Epidermal Growth Factor and Membrane Trafficking: Egf Receptor Activation of Endocytosis Requires Rab5a
title_fullStr Epidermal Growth Factor and Membrane Trafficking: Egf Receptor Activation of Endocytosis Requires Rab5a
title_full_unstemmed Epidermal Growth Factor and Membrane Trafficking: Egf Receptor Activation of Endocytosis Requires Rab5a
title_short Epidermal Growth Factor and Membrane Trafficking: Egf Receptor Activation of Endocytosis Requires Rab5a
title_sort epidermal growth factor and membrane trafficking: egf receptor activation of endocytosis requires rab5a
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2185585/
https://www.ncbi.nlm.nih.gov/pubmed/11062256
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