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SNARE proteins mediate fusion between cytosolic lipid droplets and are implicated in insulin sensitivity

Nature Cell Biology volume 9, pages 1286–1293 (2007)Cite this article

Abstract

The accumulation of cytosolic lipid droplets in muscle and liver cells has been linked to the development of insulin resistance and type 2 diabetes1. Such droplets are formed as small structures2 that increase in size through fusion3, a process that is dependent on intact microtubules and the motor protein dynein3,4. Approximately 15% of all droplets are involved in fusion processes at a given time3. Here, we show that lipid droplets are associated with proteins involved in fusion processes in the cell: NSF (N-ethylmaleimide-sensitive-factor), α-SNAP (soluble NSF attachment protein) and the SNAREs (SNAP receptors), SNAP23 (synaptosomal-associated protein of 23 kDa), syntaxin-5 and VAMP4 (vesicle-associated membrane protein 4). Knockdown of the genes for SNAP23, syntaxin-5 or VAMP4, or microinjection of a dominant-negative mutant of α-SNAP, decreases the rate of fusion and the size of the lipid droplets. Thus, the SNARE system seems to have an important role in lipid droplet fusion. We also show that oleic acid treatment decreases the insulin sensitivity of heart muscle cells, and this sensitivity is completely restored by transfection with SNAP23. Thus, SNAP23 might be a link between insulin sensitivity and the inflow of fatty acids to the cell.

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Figure 1: Localization of SNAREs, α-SNAP and NSF on lipid droplets from NIH 3T3 cells cultured in the presence of oleic acid for 24 h.
Figure 2: SNAP23, syntaxin-5 and VAMP4 form a complex that is recognized by α-SNAP in NIH-3T3 cells.
Figure 3: Knockdown of SNAP23, syntaxin-5 or VAMP4 or microinjection with a dominant-negative mutant of α-SNAP decreases the lipid droplet volume in NIH 3T3 cells.
Figure 4: Incubation of HL-1 cells with oleic acid (OA) increases the pool of lipid droplets, decreases the plasma-membrane pool of SNAP23 and increases the pool of lipid-droplet-associated SNAP23.
Figure 5: Incubation of HL-1 cells with oleic acid (OA) results in insulin resistance that can be rescued by transfection with SNAP23.

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Acknowledgements

We thank R.S. Perkins for expert editing of the manuscript, H.Y. Gaisano for the C-terminal truncated SNAP23 and J.C. Hay for the antibodies against Sec22 and Rbet1. The work was supported by grants from the Swedish Research Council, the Swedish Foundation for Strategic Research, the Swedish Heart and Lung Foundation, the NovoNordic Foundation and the Swedish Diabetes Foundation.

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Authors and Affiliations

  1. Sahlgrenska Center for Cardiovascular and Metabolic Research, Wallenberg Laboratory, Sahlgrenska Univeristy Hospital, SE-413 45 Göteborg, Sweden.,

    Pontus Boström, Linda Andersson, Mikael Rutberg, Jeanna Perman, Ulf Lidberg, Johanna Ericson, Jan Borén & Sven-Olof Olofsson

  2. Department of Biomedicine, University of Göteborg, Göteborg, Sweden.,

    Bengt R. Johansson, Julia Fernandez-Rodriguez & Tommy Nilsson

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Correspondence to Sven-Olof Olofsson.

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Supplementary Figures S1, S2, S3, S4, S5, S6 and Supplementary Methods (PDF 6739 kb)

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Boström, P., Andersson, L., Rutberg, M. et al. SNARE proteins mediate fusion between cytosolic lipid droplets and are implicated in insulin sensitivity. Nat Cell Biol 9, 1286–1293 (2007). https://doi.org/10.1038/ncb1648

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