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Human VPS34 is required for internal vesicle formation within multivesicular endosomes

After internalization from the plasma membrane, activated EGF receptors (EGFRs) are delivered to multivesicular bodies (MVBs). Within MVBs, EGFRs are removed from the perimeter membrane to internal vesicles, thereby being sorted from transferrin receptors, which recycle back to the plasma membrane....

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Detalles Bibliográficos
Autores principales: Futter, C.E., Collinson, L.M., Backer, J.M., Hopkins, C.R.
Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 2001
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2199316/
https://www.ncbi.nlm.nih.gov/pubmed/11756475
http://dx.doi.org/10.1083/jcb.200108152
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author Futter, C.E.
Collinson, L.M.
Backer, J.M.
Hopkins, C.R.
author_facet Futter, C.E.
Collinson, L.M.
Backer, J.M.
Hopkins, C.R.
author_sort Futter, C.E.
collection PubMed
description After internalization from the plasma membrane, activated EGF receptors (EGFRs) are delivered to multivesicular bodies (MVBs). Within MVBs, EGFRs are removed from the perimeter membrane to internal vesicles, thereby being sorted from transferrin receptors, which recycle back to the plasma membrane. The phosphatidylinositol (PI) 3′-kinase inhibitor, wortmannin, inhibits internal vesicle formation within MVBs and causes EGFRs to remain in clusters on the perimeter membrane. Microinjection of isotype-specific inhibitory antibodies demonstrates that the PI 3′-kinase required for internal vesicle formation is hVPS34. In the presence of wortmannin, EGFRs continue to be delivered to lysosomes, showing that their removal from the recycling pathway and their delivery to lysosomes does not depend on inward vesiculation. We showed previously that tyrosine kinase-negative EGFRs fail to accumulate on internal vesicles of MVBs but are recycled rather than delivered to lysosomes. Therefore, we conclude that selection of EGFRs for inclusion on internal vesicles requires tyrosine kinase but not PI 3′-kinase activity, whereas vesicle formation requires PI 3′-kinase activity. Finally, in wortmannin-treated cells there is increased EGF-stimulated tyrosine phosphorylation when EGFRs are retained on the perimeter membrane of MVBs. Therefore, we suggest that inward vesiculation is involved directly with attenuating signal transduction.
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spelling pubmed-21993162008-05-01 Human VPS34 is required for internal vesicle formation within multivesicular endosomes Futter, C.E. Collinson, L.M. Backer, J.M. Hopkins, C.R. J Cell Biol Article After internalization from the plasma membrane, activated EGF receptors (EGFRs) are delivered to multivesicular bodies (MVBs). Within MVBs, EGFRs are removed from the perimeter membrane to internal vesicles, thereby being sorted from transferrin receptors, which recycle back to the plasma membrane. The phosphatidylinositol (PI) 3′-kinase inhibitor, wortmannin, inhibits internal vesicle formation within MVBs and causes EGFRs to remain in clusters on the perimeter membrane. Microinjection of isotype-specific inhibitory antibodies demonstrates that the PI 3′-kinase required for internal vesicle formation is hVPS34. In the presence of wortmannin, EGFRs continue to be delivered to lysosomes, showing that their removal from the recycling pathway and their delivery to lysosomes does not depend on inward vesiculation. We showed previously that tyrosine kinase-negative EGFRs fail to accumulate on internal vesicles of MVBs but are recycled rather than delivered to lysosomes. Therefore, we conclude that selection of EGFRs for inclusion on internal vesicles requires tyrosine kinase but not PI 3′-kinase activity, whereas vesicle formation requires PI 3′-kinase activity. Finally, in wortmannin-treated cells there is increased EGF-stimulated tyrosine phosphorylation when EGFRs are retained on the perimeter membrane of MVBs. Therefore, we suggest that inward vesiculation is involved directly with attenuating signal transduction. The Rockefeller University Press 2001-12-24 /pmc/articles/PMC2199316/ /pubmed/11756475 http://dx.doi.org/10.1083/jcb.200108152 Text en Copyright © 2001, 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 Article
Futter, C.E.
Collinson, L.M.
Backer, J.M.
Hopkins, C.R.
Human VPS34 is required for internal vesicle formation within multivesicular endosomes
title Human VPS34 is required for internal vesicle formation within multivesicular endosomes
title_full Human VPS34 is required for internal vesicle formation within multivesicular endosomes
title_fullStr Human VPS34 is required for internal vesicle formation within multivesicular endosomes
title_full_unstemmed Human VPS34 is required for internal vesicle formation within multivesicular endosomes
title_short Human VPS34 is required for internal vesicle formation within multivesicular endosomes
title_sort human vps34 is required for internal vesicle formation within multivesicular endosomes
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2199316/
https://www.ncbi.nlm.nih.gov/pubmed/11756475
http://dx.doi.org/10.1083/jcb.200108152
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