Phospholipase D Stimulates Release of Nascent Secretory Vesicles from the trans-Golgi Network
Ye-Guang Chen,Anirban Siddhanta,Cary D. Austin,Scott M. Hammond,Tsung Chang Sung,Michael A. Frohman,Andrew J. Morris,Dennis Shields +7 more
TLDR
It is demonstrated that immunoaffinity-purified human PLD1 stimulated nascent secretory vesicle budding from the TGN and ARF-1 stimulated endogenous PLD activity in Golgi membranes approximately threefold and this activation correlated with its enhancement of vesicles budding.Abstract:
Phospholipase D (PLD) is a phospholipid hydrolyzing enzyme whose activation has been implicated in mediating signal transduction pathways, cell growth, and membrane trafficking in mammalian cells. Several laboratories have demonstrated that small GTP-binding proteins including ADP-ribosylation factor (ARF) can stimulate PLD activity in vitro and an ARF-activated PLD activity has been found in Golgi membranes. Since ARF-1 has also been shown to enhance release of nascent secretory vesicles from the TGN of endocrine cells, we hypothesized that this reaction occurred via PLD activation. Using a permeabilized cell system derived from growth hormone and prolactin-secreting pituitary GH3 cells, we demonstrate that immunoaffinity-purified human PLD1 stimulated nascent secretory vesicle budding from the TGN approximately twofold. In contrast, a similarly purified but enzymatically inactive mutant form of PLD1, designated Lys898Arg, had no effect on vesicle budding when added to the permeabilized cells. The release of nascent secretory vesicles from the TGN was sensitive to 1% 1-butanol, a concentration that inhibited PLD-catalyzed formation of phosphatidic acid. Furthermore, ARF-1 stimulated endogenous PLD activity in Golgi membranes approximately threefold and this activation correlated with its enhancement of vesicle budding. Our results suggest that ARF regulation of PLD activity plays an important role in the release of nascent secretory vesicles from the TGN.read more
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Interaction of the Type Iα PIPkinase with phospholipase D: a role for the local generation of phosphatidylinositol 4,5-bisphosphate in the regulation of PLD2 activity
Nullin Divecha,Mieke Roefs,Jonathan R. Halstead,Sabine d'Andrea,Mar Fernandez‐Borga,L. C. J. M. Oomen,Kahlid M. Saqib,Michael J.O. Wakelam,Clive D'Santos +8 more
TL;DR: It is shown that the physiological requirement of PLD enzymes for PtdIns(4,5)P2 is critical and that PLD2 activity in vivo can be regulated solely by the levels of this key intracellular lipid.
Journal ArticleDOI
Phospholipase D structure and regulation.
TL;DR: In vivo roles for phospholipase D suggest that it may be important for multiples steps in regulated secretion and membrane biogenesis.
Journal ArticleDOI
PLD2 Complexes with the EGF Receptor and Undergoes Tyrosine Phosphorylation at a Single Site upon Agonist Stimulation
TL;DR: It is demonstrated here for the first time agonist-stimulated activation of both PLD1 and PLD2 in vivo and evidence of a distinct type of interaction for each isoform with the EGF receptor is provided.
Journal ArticleDOI
Inhibition of phosphatidic acid synthesis alters the structure of the Golgi apparatus and inhibits secretion in endocrine cells.
TL;DR: The results suggest that PA stimulation of PtdIns(4,5)P2 synthesis is required for maintaining the structural integrity and function of the Golgi apparatus.
Journal ArticleDOI
Phospholipase D signaling: orchestration by PIP2 and small GTPases
TL;DR: This review highlights the regulation ofPLD by membrane receptors, and describes how the close encounter of PLD and PIP5K isoforms with small GTPases permits the execution of specific cellular functions.
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ADP-ribosylation factor, a small GTP-dependent regulatory protein, stimulates phospholipase D activity
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