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Vol. 15, Issue 1, 99-110, January 2004
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* Institute for Molecular Bioscience, Centre for Microscopy and Microanalysis, and School of Biomedical Sciences, University of Queensland, Queensland 4072, Australia;
Departament de Biologia Cel·lular, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Facultat de Medicina, Universitat de Barcelona, Barcelona 08036, Spain
Submitted June 6, 2003;
Revised July 18, 2003;
Accepted September 3, 2003
Monitoring Editor: Jean Gruenberg
Caveolins are a crucial component of caveolae but have also been localized to the Golgi complex, and, under some experimental conditions, to lipid bodies (LBs). The physiological relevance and dynamics of LB association remain unclear. We now show that endogenous caveolin-1 and caveolin-2 redistribute to LBs in lipid loaded A431 and FRT cells. Association with LBs is regulated and reversible; removal of fatty acids causes caveolin to rapidly leave the lipid body. We also show by subcellular fractionation, light and electron microscopy that during the first hours of liver regeneration, caveolins show a dramatic redistribution from the cell surface to the newly formed LBs. At later stages of the regeneration process (when LBs are still abundant), the levels of caveolins in LBs decrease dramatically. As a model system to study association of caveolins with LBs we have used brefeldin A (BFA). BFA causes rapid redistribution of endogenous caveolins to LBs and this association was reversed upon BFA washout. Finally, we have used a dominant negative LB-associated caveolin mutant (cavDGV) to study LB formation and to examine its effect on LB function. We now show that the cavDGV mutant inhibits microtubule-dependent LB motility and blocks the reversal of lipid accumulation in LBs.
Abbreviations used: BFA, brefeldin A; BHK, baby hamster kidney cells; CavDGV, caveolin 3 DGV truncation; CyHx, cyclohexamide; DAPI, 4',6-diamidino-2-phenylindole dihydrochlorid; ER, endoplasmic reticulum; FRT, Fisher rat tyroid cells; GFP, green fluorescent protein; LB, lipid body; OlAc, oleic acid; PM, plasma membrane; PH, partial hepatectomy; YFP, yellow fluorescent protein.
Online version of this article contains video material for some figures. Online version is available at www.molbiolcell.org.
Corresponding author. E-mail address: R.Parton{at}imb.uq.edu.au.
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