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Papers In Press, published online ahead of print May 10, 2004
Department of Life Science, University of Hyogo, Kamigori, Hyogo 678-1297
Corresponding Author: osumi{at}sci.u-hyogo.ac.jp
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. Over-expressed 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.
J. Biol. Chem, 10.1074/jbc.M403920200
Submitted on April 8, 2004
Revised on May 7, 2004
Accepted on May 10, 2004
CGI-58 interacts with perilipin and is localized to lipid droplets: Possible involvement of CGI-58 mislocalization in chanarin-Dorfman syndrome
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