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Functional genomic screen reveals genes involved in lipid-droplet formation and utilization

Abstract

Eukaryotic cells store neutral lipids in cytoplasmic lipid droplets1,2 enclosed in a monolayer of phospholipids and associated proteins3,4. These dynamic organelles5 serve as the principal reservoirs for storing cellular energy and for the building blocks for membrane lipids. Excessive lipid accumulation in cells is a central feature of obesity, diabetes and atherosclerosis, yet remarkably little is known about lipid-droplet cell biology. Here we show, by means of a genome-wide RNA interference (RNAi) screen in Drosophila S2 cells that about 1.5% of all genes function in lipid-droplet formation and regulation. The phenotypes of the gene knockdowns sorted into five distinct phenotypic classes. Genes encoding enzymes of phospholipid biosynthesis proved to be determinants of lipid-droplet size and number, suggesting that the phospholipid composition of the monolayer profoundly affects droplet morphology and lipid utilization. A subset of the Arf1–COPI vesicular transport proteins also regulated droplet morphology and lipid utilization, thereby identifying a previously unrecognized function for this machinery. These phenotypes are conserved in mammalian cells, suggesting that insights from these studies are likely to be central to our understanding of human diseases involving excessive lipid storage.

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Figure 1: Oleate increases the formation of lipid droplets in Drosophila S2 cells.
Figure 2: Genome-wide screen identified genes regulating the formation of lipid droplets.
Figure 3: Phosphatidylcholine content regulates the size and abundance of lipid droplets.
Figure 4: ArfI–COPI complex members function in the formation of lipid droplets.

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Acknowledgements

We thank I. Vasenkova and R. De Breuil for help with dsRNA synthesis; K. Warner for help with yeast work; M. Schuldiner, E. Griffis, T. Fazzio, E. Herker, S. Stymne, B. Panning and M. Ott for reagents; D. B. Jones and G. Howard for assistance with manuscript preparation; G. Schoenhofer for web access for the database; members of the Farese, Vale and Walter laboratories for discussions; and D. Srivastava and S. Yamanaka for critical reading of the manuscript. This work was supported by a Freedom to Discover Award from Bristol–Myers Squibb and National Institutes of Health grant R21 DK078254-01 (to R.F.), a David and Mary Phillips postdoctoral fellowship award (to Y.G.), the Human Frontier Science Program Organization (T.C.W.), the Howard Hughes Medical Institute (P.W. and R.D.V.) and the J. David Gladstone Institutes.

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Correspondence to Tobias C. Walther or Robert V. Farese.

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The file contains Supplementary Methods, Supplementary Tables S1-S3 and Supplementary Figures S1-S5. (PDF 4198 kb)

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Guo, Y., Walther, T., Rao, M. et al. Functional genomic screen reveals genes involved in lipid-droplet formation and utilization. Nature 453, 657–661 (2008). https://doi.org/10.1038/nature06928

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