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J. Biol. Chem., Vol. 282, Issue 30, 21829-21837, July 27, 2007

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Active ADP-ribosylation Factor-1 (ARF1) Is Required for Mitotic Golgi Fragmentation^*

Yi Xiang, Joachim Seemann, Blaine Bisel, Sukanya Punthambaker, and Yanzhuang Wang¹

From the Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, Michigan 48109-1048 and the Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, Texas 75390-9039

In mammalian cells the Golgi apparatus undergoes an extensive disassembly process at the onset of mitosis that is believed to facilitate equal partitioning of this organelle into the two daughter cells. However, the underlying mechanisms for this fragmentation process are so far unclear. Here we have investigated the role of the ADP-ribosylation factor-1 (ARF1) in this process to determine whether Golgi fragmentation in mitosis is mediated by vesicle budding. ARF1 is a small GTPase that is required for COPI vesicle formation from the Golgi membranes. Treatment of Golgi membranes with mitotic cytosol or with purified coatomer together with wild type ARF1 or its constitutive active form, but not the inactive mutant, converted the Golgi membranes into COPI vesicles. ARF1-depleted mitotic cytosol failed to fragment Golgi membranes. ARF1 is associated with Golgi vesicles generated in vitro and with vesicles in mitotic cells. In addition, microinjection of constitutive active ARF1 did not affect mitotic Golgi fragmentation or cell progression through mitosis. Our results show that ARF1 is active during mitosis and that this activity is required for mitotic Golgi fragmentation.

Received for publication, December 21, 2006 , and in revised form, June 11, 2007.

^* 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. 1–3.

¹ To whom correspondence should be addressed: Dept. of Molecular, Cellular and Developmental Biology, University of Michigan, 830 North University Ave., Ann Arbor, MI 48109-1048. Tel.: 734-936-2134; E-mail: yzwang{at}umich.edu.

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				D. Tang, K. Mar, G. Warren, and Y. Wang Molecular Mechanism of Mitotic Golgi Disassembly and Reassembly Revealed by a Defined Reconstitution Assay J. Biol. Chem., March 7, 2008; 283(10): 6085 - 6094. [Abstract] [Full Text] [PDF]