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J. Biol. Chem., Vol. 282, Issue 47, 34085-34092, November 23, 2007

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Requirement of Phospholipase D for Ilimaquinone-induced Golgi Membrane Fragmentation^*

From the Division of Biochemistry, Department of Molecular and Cellular Biology, Kobe University Graduate School of Medicine, Kobe 650-0017, Japan

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 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 2-butanol, 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.

Received for publication, July 9, 2007 , and in revised form, August 17, 2007.

^* This work was supported in part by a grant-in-aid for Center of Excellence Research from the Ministry of Education, Science, Sports, and Culture of Japan. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains supplemental Figs. S1–S3.

¹ To whom correspondence should be addressed. Tel.: 81-78-382-5420; Fax: 81-78-382-5439; E-mail: snakamur{at}kobe-u.ac.jp.

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