Regulation of phospholipase D1 subcellular cycling through coordination of multiple membrane association motifs.
Fiche publication
Date publication
juillet 2003
Journal
The Journal of cell biology
Auteurs
Membres identifiés du Cancéropôle Est :
Dr BADER Marie-France, Dr CHASSEROT-GOLAZ Sylvette, Dr VITALE Nicolas
Tous les auteurs :
Du G, Altshuller YM, Vitale N, Huang P, Chasserot-Golaz S, Morris AJ, Bader MF, Frohman MA
Lien Pubmed
Résumé
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 PI4,5P2-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 PI4,5P2-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.
Mots clés
Amino Acid Motifs, Animals, COS Cells, Cell Membrane, enzymology, Cercopithecus aethiops, Endosomes, enzymology, Enzyme Activation, Exocytosis, Gene Expression Regulation, Enzymologic, Golgi Apparatus, enzymology, Membrane Microdomains, enzymology, Models, Biological, Mutagenesis, Site-Directed, PC12 Cells, Phosphatidylinositol Phosphates, metabolism, Phospholipase D, chemistry, Rats, Subcellular Fractions, enzymology
Référence
J. Cell Biol.. 2003 Jul;162(2):305-15