Requirement of Phospholipase D for Ilimaquinone-induced Golgi Membrane Fragmentation
Journal of Biological Chemistry
Although organelles such as the endoplasmic reticulum and Golgi apparatus are highly compartmentalized, these organelles are interconnected through a network of vesicular trafficking. The marine sponge metabolite ilimaquinone (IQ) is known to induce Golgi membrane fragmentation and is widely used to study the mechanism of vesicular trafficking. Although IQ treatment causes protein kinase D (PKD) activation, the detailed mechanism of IQ-induced Golgi membrane fragmentation remains unclear. In
... ains unclear. In this work, we found that IQ treatment of cells caused a robust activation of phospholipase D (PLD). In the presence of 1-butanol but not 2butanol, IQ-induced Golgi membrane fragmentation was completely blocked. In addition, IQ failed to induce Golgi membrane fragmentation in PLD knock-out DT40 cells. Furthermore, IQ-induced PKD activation was completely blocked by treatment with either 1-butanol or propranolol. Notably, IQ-induced Golgi membrane fragmentation was also blocked by propranolol treatment. These results indicate that PLD-catalyzed formation of phosphatidic acid is a prerequisite for IQ-induced Golgi membrane fragmentation and that enzymatic conversion of phosphatidic acid to diacylglycerol is necessary for subsequent activation of PKD and IQ-induced Golgi membrane fragmentation. .03% Me 2 SO) or with various concentrations of IQ. As a positive control PLD was activated for 1 h with 100 nM PMA. PLD activity is expressed as a percentage of the radioactive [ 14 C]phosphatidylbutanol (PtdBut) in the total radioactivity found in all spots in a particular lane. Data are the means Ϯ S.E. of three independent experiments carried out in triplicate.