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Vol. 18, Issue 9, 3250-3263, September 2007
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*Departament de Biologia Cellular i Anatomia Patològica, Facultat de Medicina and Institut d'Investigacions Biomèdiques August Pi i Sunyer, and Institut de Nanociència i Nanotecnologia, Universitat de Barcelona, 08036 Barcelona, Spain; Cellular Architecture and Dynamics and ¶Biochemical Physiology, Science Faculty and Institute of Biomembranes, Utrecht University, 3584 CH Utrecht, The Netherlands; and Institut de Neurociències i Departament de Bioquímica i Biologia Molecular, Facultat de Medicina, Universitat Autònoma de Barcelona, 08193 Bellaterra, Barcelona, Spain; and ||Departamento de Inmunología y Oncología, Instituto Nacional de Biotecnología, Universidad Autónoma de Madrid-Consejo Superior de Investigaciones Científicas, 28049 Madrid, Spain
Submitted April 12, 2007;
Revised May 30, 2007;
Accepted June 6, 2007
Monitoring Editor: Vivek Malhotra
Diacylglycerol is necessary for trans-Golgi network (TGN) to cell surface transport, but its functional relevance in the early secretory pathway is unclear. Although depletion of diacylglycerol did not affect ER-to-Golgi transport, it led to a redistribution of the KDEL receptor to the Golgi, indicating that Golgi-to-ER transport was perturbed. Electron microscopy revealed an accumulation of COPI-coated membrane profiles close to the Golgi cisternae. Electron tomography showed that the majority of these membrane profiles originate from coated buds, indicating a block in membrane fission. Under these conditions the Golgi-associated pool of ARFGAP1 was reduced, but there was no effect on the binding of coatomer or the membrane fission protein CtBP3/BARS to the Golgi. The addition of 1,2-dioctanoyl-sn-glycerol or the diacylglycerol analogue phorbol 12,13-dibutyrate reversed the effects of endogenous diacylglycerol depletion. Our findings implicate diacylglycerol in the retrograde transport of proteins from Golgi to the ER and suggest that it plays a critical role at a late stage of COPI vesicle formation.
The online version of this article contains supplemental material at MBC Online (http://www.molbiolcell.org).
Address correspondence to: Gustavo Egea (gegea{at}ub.edu).
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