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First published online 24 May 2005
doi: 10.1242/jcs.02401

Journal of Cell Science 118, 2601-2611 (2005)
Published by The Company of Biologists 2005
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Research Article

Rab18 localizes to lipid droplets and induces their close apposition to the endoplasmic reticulum-derived membrane

Shintaro Ozeki1,*, Jinglei Cheng1,*, Kumi Tauchi-Sato1, Naoya Hatano2, Hisaaki Taniguchi2,3 and Toyoshi Fujimoto1,{ddagger}

1 Department of Anatomy and Molecular Cell Biology, Graduate School of Medicine, Nagoya University, 65 Tsurumai, Showa, Nagoya 466-8550, Japan
2 Harima Institute at SPring-8, RIKEN, Mikazuki, Sayo, Hyogo 679-5148, Japan
3 Institute for Enzyme Research, The University of Tokushima, Tokushima 770-8503, Japan

{ddagger} Author for correspondence (e-mail: tfujimot{at}med.nagoya-u.ac.jp)

Accepted 17 March 2005

Lipid droplets (LDs) are organelles that store neutral lipids, but their regulatory mechanism is not well understood. In the present study, we identified Rab18 as an LD component of HepG2 cells by proteomic analysis, and confirmed its localization by immunohistochemistry and western blotting. Wild-type and dominant-active Rab18 localized to LDs but the dominant-negative form did not. Endogenous Rab18 coexisted with adipocyte differentiation-related protein (ADRP) in LDs, but the labeling intensity of the two proteins showed clear reciprocity. Consistent with this observation, overexpression of Rab18 induced a decrease in the amounts of ADRP in LDs in HepG2 and BALB/c 3T3 cells. Furthermore, Rab18 overexpression caused close apposition of LDs to membrane cisternae connected to the rough ER. Two other procedures that decrease ADRP, i.e. RNA interference and brefeldin A treatment, induced the same morphological change, indicating that decrease in ADRP was the cause of the LD-ER apposition. In accordance with similar structures found between ER and other organelles, we propose that the ER membrane apposed to LDs should be named the LD-associated membrane, or LAM. The present results suggested that Rab18 regulates LAM formation, which is likely to be involved in mobilizing lipid esters stored in LDs.

Key words: Lipid droplet, Mass spectrometry, Rab18, Endoplasmic reticulum, Membrane apposition




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