Phospholipase D2 is localized to the rims of the Golgi apparatus in mammalian cells.
Phospholipase D (PLD) hydrolyzes phosphatidylcholine to generate phosphatidic acid, a molecule known to have multiple physiological roles, including release of nascent secretory vesicles from the trans-Golgi network. In mammalian cells two forms of the enzyme, PLD1 and PLD2, have been described. We recently demonstrated that PLD1 is localized to the Golgi apparatus, nuclei, and to a lesser extent, plasma membrane. Due to its low abundance, the intracellular localization of PLD2 has been characterized only indirectly through overexpression of chimeric proteins. Using antibodies specific to PLD2, together with immunofluorescence microscopy, herein we demonstrate that a significant fraction of endogenous PLD2 localized to the perinuclear Golgi region and was also distributed throughout cells in dense cytoplasmic puncta; a fraction of which colocalized with caveolin-1 and the plasma membrane. On treatment with brefeldin A, PLD2 translocated into the nucleus in a manner similar to PLD1, suggesting a potential role in nuclear signaling. Most significantly, cryoimmunogold electron microscopy demonstrated that in pituitary GH(3) cells >90% of PLD2 present in the Golgi apparatus was localized to cisternal rims and peri-Golgi vesicles exclusively. The data are consistent with a model whereby PLD2 plays a role in Golgi vesicular transport.
Freyberg
Zachary
Z
Department of Developmental and Molecular Biology, Albert Einstein College of Medicine, Bronx, New York 10461, USA.
Bourgoin
Sylvain
S
Shields
Dennis
D
D000818:Animals; D000970:Antineoplastic Agents / Q000378:metabolism; D015415:Biomarkers; D020126:Brefeldin A / Q000378:metabolism; D002460:Cell Line; D020285:Cryoelectron Microscopy; D006056:Golgi Apparatus / Q000201:enzymology*; D007150:Immunohistochemistry; D007527:Isoenzymes / Q000378:metabolism; D007668:Kidney / Q000166:cytology / Q000201:enzymology; D016253:Microscopy, Immunoelectron; D015739:Nocodazole / Q000378:metabolism; D010739:Phospholipase D / Q000378:metabolism*; D011500:Protein Synthesis Inhibitors / Q000378:metabolism; D021381:Protein Transport / Q000502:physiology; D051381:Rats; D015398:Signal Transduction / Q000502:physiology; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; D015415:Biomarkers; D007527:Isoenzymes; D011500:Protein Synthesis Inhibitors; D020126:Brefeldin A; C105606:phospholipase D2; D010739:Phospholipase D; C106801:phospholipase D1; D015739:Nocodazole