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MBC in Press, published online ahead of print June 13, 2007
Mol. Biol. Cell 10.1091/mbc.E07-04-0334

A more recent version of this article appeared on September 1, 2007
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Submitted on April 12, 2007
Revised on May 30, 2007
Accepted on June 6, 2007

Diacylglycerol Is Required for the Formation of COPI Vesicles in the Golgi-to-ER Transport Pathway

Inés Fernández-Ulibarri,* Montserrat Vilella,* Francisco Lázaro-Diéguez,*{dagger} Elisabet Sarri,* Susana E. Martínez,* Nuria Jiménez,{ddagger} Enrique Claro,{sect} Isabel Mérida,|| Koert N.J. Burger,¶ and Gustavo Egea*{dagger}

*Departament de Biologia Cellular i Anatomia Patològica, Facultat de Medicina and Institut d’Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), and {dagger}Institut de Nanociència i Nanotecnologia (IN2UB), Universitat de Barcelona, 08036 Barcelona, Spain; {ddagger}Cellular Architecture and Dynamics and Biochemical Physiology, Science Faculty and Institute of Biomembranes, Utrecht University, 3584 CH Utrecht, The Netherlands; {sect}Institut de Neurociències i Departament de Bioquímica i Biologia Molecular, Facultat de Medicina, UAB, 08193 Bellaterra, Barcelona, Spain; ||Departamento de Inmunología y Oncología, Instituto Nacional de Biotecnología, UAM-CSIC, 28049 Madrid, Spain

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.

Address correspondence to: Gustavo Egea (gegea{at}ub.edu)




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