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Originally published In Press as doi:10.1074/jbc.M403920200 on May 10, 2004
J. Biol. Chem., Vol. 279, Issue 29, 30490-30497, July 16, 2004
CGI-58 Interacts with Perilipin and Is Localized to Lipid Droplets
POSSIBLE INVOLVEMENT OF CGI-58 MISLOCALIZATION IN CHANARIN-DORFMAN SYNDROME*
Tomohiro Yamaguchi,
Naoto Omatsu,
Shuhei Matsushita, and
Takashi Osumi
From the
Graduate School of Life Science, Himeji Institute of Technology, University of Hyogo, 3-2-1 Koto, Kamigori, Hyogo 678-1297, Japan
Lipid droplets (LDs) are a class of ubiquitous cellular organelles that are involved in lipid storage and metabolism. Although the mechanisms of the biogenesis of LDs are still unclear, a set of proteins called the PAT domain family have been characterized as factors associating with LDs. Perilipin, a member of this family, is expressed exclusively in the adipose tissue and regulates the breakdown of triacylglycerol in LDs via its phosphorylation. In this study, we used a yeast two-hybrid system to examine the potential function of perilipin. We found direct interaction between perilipin and CGI-58, a deficiency of which correlated with the pathogenesis of Chanarin-Dorfman syndrome (CDS). Endogenous CGI-58 was distributed predominantly on the surface of LDs in differentiated 3T3-L1 cells, and its expression increased during adipocyte differentiation. Overexpressed CGI-58 tagged with GFP gathered at the surface of LDs and colocalized with perilipin. This interaction seems physiologically important because CGI-58 mutants carrying an amino acid substitution identical to that found in CDS lost the ability to be recruited to LDs. These mutations significantly weakened the binding of CGI-58 with perilipin, indicating that the loss of this interaction is involved in the etiology of CDS. Furthermore, we identified CGI-58 as a binding partner of ADRP, another PAT domain protein expressed ubiquitously, by yeast two-hybrid assay. GFP-CGI-58 expressed in non-differentiated 3T3-L1 or CHO-K1 cells was colocalized with ADRP, and the CGI-58 mutants were not recruited to LDs carrying ADRP, indicating that CGI-58 may also cooperate with ADRP.
Received for publication, April 8, 2004
, and in revised form, May 7, 2004.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EBI Data Bank with accession number(s) AY550934.
* This work was supported by grants from Uehara Memorial Foundation (to T. Y.), a grant-in-aid for Scientific Research from the Japan Society for the Promotion of Science (to T. O.), and the 21st Century Center of Excellence Program. 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.
To whom correspondence should be addressed. Tel.: 81-791-58-0194; Fax: 81-791-58-0193; E-mail: osumi{at}sci.u-hyogo.ac.jp.

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Copyright © 2004 by the American Society for Biochemistry and Molecular Biology.
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