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Regulation of phospholipase D1 subcellular cycling through coordination of multiple membrane association motifs
The signaling enzyme phospholipase D1 (PLD1) facilitates membrane vesicle trafficking. Here, we explore how PLD1 subcellular localization is regulated via Phox homology (PX) and pleckstrin homology (PH) domains and a PI4,5P(2)-binding site critical for its activation. PLD1 localized to perinuclear e...
Autores principales: | , , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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The Rockefeller University Press
2003
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2172799/ https://www.ncbi.nlm.nih.gov/pubmed/12876278 http://dx.doi.org/10.1083/jcb.200302033 |
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author | Du, Guangwei Altshuller, Yelena M. Vitale, Nicolas Huang, Ping Chasserot-Golaz, Sylvette Morris, Andrew J. Bader, Marie-France Frohman, Michael A. |
author_facet | Du, Guangwei Altshuller, Yelena M. Vitale, Nicolas Huang, Ping Chasserot-Golaz, Sylvette Morris, Andrew J. Bader, Marie-France Frohman, Michael A. |
author_sort | Du, Guangwei |
collection | PubMed |
description | The signaling enzyme phospholipase D1 (PLD1) facilitates membrane vesicle trafficking. Here, we explore how PLD1 subcellular localization is regulated via Phox homology (PX) and pleckstrin homology (PH) domains and a PI4,5P(2)-binding site critical for its activation. PLD1 localized to perinuclear endosomes and Golgi in COS-7 cells, but on cellular stimulation, translocated to the plasma membrane in an activity-facilitated manner and then returned to the endosomes. The PI4,5P(2)-interacting site sufficed to mediate outward translocation and association with the plasma membrane. However, in the absence of PX and PH domains, PLD1 was unable to return efficiently to the endosomes. The PX and PH domains appear to facilitate internalization at different steps. The PH domain drives PLD1 entry into lipid rafts, which we show to be a step critical for internalization. In contrast, the PX domain appears to mediate binding to PI5P, a lipid newly recognized to accumulate in endocytosing vesicles. Finally, we show that the PH domain–dependent translocation step, but not the PX domain, is required for PLD1 to function in regulated exocytosis in PC12 cells. We propose that PLD1 localization and function involves regulated and continual cycling through a succession of subcellular sites, mediated by successive combinations of membrane association interactions. |
format | Text |
id | pubmed-2172799 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2003 |
publisher | The Rockefeller University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-21727992008-05-01 Regulation of phospholipase D1 subcellular cycling through coordination of multiple membrane association motifs Du, Guangwei Altshuller, Yelena M. Vitale, Nicolas Huang, Ping Chasserot-Golaz, Sylvette Morris, Andrew J. Bader, Marie-France Frohman, Michael A. J Cell Biol Article The signaling enzyme phospholipase D1 (PLD1) facilitates membrane vesicle trafficking. Here, we explore how PLD1 subcellular localization is regulated via Phox homology (PX) and pleckstrin homology (PH) domains and a PI4,5P(2)-binding site critical for its activation. PLD1 localized to perinuclear endosomes and Golgi in COS-7 cells, but on cellular stimulation, translocated to the plasma membrane in an activity-facilitated manner and then returned to the endosomes. The PI4,5P(2)-interacting site sufficed to mediate outward translocation and association with the plasma membrane. However, in the absence of PX and PH domains, PLD1 was unable to return efficiently to the endosomes. The PX and PH domains appear to facilitate internalization at different steps. The PH domain drives PLD1 entry into lipid rafts, which we show to be a step critical for internalization. In contrast, the PX domain appears to mediate binding to PI5P, a lipid newly recognized to accumulate in endocytosing vesicles. Finally, we show that the PH domain–dependent translocation step, but not the PX domain, is required for PLD1 to function in regulated exocytosis in PC12 cells. We propose that PLD1 localization and function involves regulated and continual cycling through a succession of subcellular sites, mediated by successive combinations of membrane association interactions. The Rockefeller University Press 2003-07-21 /pmc/articles/PMC2172799/ /pubmed/12876278 http://dx.doi.org/10.1083/jcb.200302033 Text en Copyright © 2003, 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 Du, Guangwei Altshuller, Yelena M. Vitale, Nicolas Huang, Ping Chasserot-Golaz, Sylvette Morris, Andrew J. Bader, Marie-France Frohman, Michael A. Regulation of phospholipase D1 subcellular cycling through coordination of multiple membrane association motifs |
title | Regulation of phospholipase D1 subcellular cycling through coordination of multiple membrane association motifs |
title_full | Regulation of phospholipase D1 subcellular cycling through coordination of multiple membrane association motifs |
title_fullStr | Regulation of phospholipase D1 subcellular cycling through coordination of multiple membrane association motifs |
title_full_unstemmed | Regulation of phospholipase D1 subcellular cycling through coordination of multiple membrane association motifs |
title_short | Regulation of phospholipase D1 subcellular cycling through coordination of multiple membrane association motifs |
title_sort | regulation of phospholipase d1 subcellular cycling through coordination of multiple membrane association motifs |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2172799/ https://www.ncbi.nlm.nih.gov/pubmed/12876278 http://dx.doi.org/10.1083/jcb.200302033 |
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