Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

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.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

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.

Similar content being viewed by others

References

  1. Taskinen, M. R. Diabetic dyslipidaemia: from basic research to clinical practice. Diabetologia 46, 733–749 (2003).

    Article  Google Scholar 

  2. Marchesan, D. et al. A phospholipase D-dependent process forms lipid droplets containing caveolin, adipocyte differentiation-related protein, and vimentin in a cell-free system. J. Biol. Chem. 278, 27293–27300 (2003).

    Article  CAS  Google Scholar 

  3. Bostrom, P. et al. Cytosolic lipid droplets increase in size by microtubule-dependent complex formation. Arterioscler. Thromb. Vasc. Biol. 25, 1945–1951 (2005).

    Article  Google Scholar 

  4. Andersson, L. et al. PLD1 and ERK2 regulate cytosolic lipid droplet formation. J. Cell Sci. 119, 2246–2257 (2006).

    Article  CAS  Google Scholar 

  5. Brown, D. A. Lipid droplets: Proteins floating on a pool of fat. Curr. Biol. 11, R446–R449 (2001).

    Article  CAS  Google Scholar 

  6. Londos, C., Sztalryd, C., Tansey, J.T. & Kimmel, A. R. Role of PAT proteins in lipid metabolism. Biochimie 87, 45–49 (2005).

    Article  CAS  Google Scholar 

  7. Martin, S. & Parton, R.G. Lipid droplets: a unified view of a dynamic organelle. Nature Rev. Mol. Cell Biol. 7, 373–378 (2006).

    Article  CAS  Google Scholar 

  8. Brasaemle, D. L., Dolios, G., Shapiro, L., & Wang, R. Proteomic analysis of proteins associated with lipid droplets of basal and lipolytically stimulated 3T3-L1 adipocytes. J. Biol. Chem. 279, 46835–46842 (2004).

    Article  CAS  Google Scholar 

  9. Liu, P. et al. Chinese hamster ovary K2 cell lipid droplets appear to be metabolic organelles involved in membrane traffic. J. Biol. Chem. 279, 3787–3792 (2004).

    Article  CAS  Google Scholar 

  10. Nakamura, N., Banno, Y. & Tamiya-Koizumi, K. Arf1-dependent PLD1 is localized to oleic acid-induced lipid droplets in NIH3T3 cells. Biochem. Biophys. Res. Commun. 335, 117–123 (2005).

    Article  CAS  Google Scholar 

  11. Bonifacino, J. S. & Glick, B. S. The mechanisms of vesicle budding and fusion. Cell 116, 153–166 (2004).

    Article  CAS  Google Scholar 

  12. Short, B. & Barr, F. A. Membrane fusion: caught in a trap. Curr. Biol. 14, R187–R189 (2004).

    Article  CAS  Google Scholar 

  13. Hong, W. SNAREs and traffic. Biochim. Biophys. Acta 1744, 493–517 (2005).

    PubMed  Google Scholar 

  14. Jahn, R. & Scheller, R. H. SNAREs—engines for membrane fusion. Nature Rev. Mol. Cell Biol. 7, 631–643 (2006).

    Article  CAS  Google Scholar 

  15. Kawanishi, M. et al. Role of SNAP23 in insulin-induced translocation of GLUT4 in 3T3-L1 adipocytes. Mediation of complex formation between syntaxin4 and VAMP2. J. Biol. Chem. 275, 8240–8247 (2000).

    Article  CAS  Google Scholar 

  16. Huang, X. et al. Cholecystokinin-regulated exocytosis in rat pancreatic acinar cells is inhibited by a C-terminus truncated mutant of SNAP-23. Pancreas 23, 125–133 (2001).

    Article  CAS  Google Scholar 

  17. High, S. & Abell, B. M. Tail-anchored protein biosynthesis at the endoplasmic reticulum: the same but different. Biochem. Soc. Trans. 32, 659–662 (2004).

    Article  CAS  Google Scholar 

  18. Bryant, N. J., Govers, R. & James, D. E. Regulated transport of the glucose transporter GLUT4. Nature Rev. Mol. Cell Biol. 3, 267–277 (2002).

    Article  CAS  Google Scholar 

  19. Ishiki, M. & Klip, A. Minireview: recent developments in the regulation of glucose transporter-4 traffic: new signals, locations, and partners. Endocrinology 146, 5071–5078 (2005).

    Article  CAS  Google Scholar 

  20. Foster, L. J., Yaworsky, K., Trimble, W. S., & Klip, A. SNAP23 promotes insulin-dependent glucose uptake in 3T3-L1 adipocytes: possible interaction with cytoskeleton. Am. J. Physiol. 276, C1108–C1114 (1999).

    Article  CAS  Google Scholar 

  21. Claycomb, W. C. et al. HL-1 cells: a cardiac muscle cell line that contracts and retains phenotypic characteristics of the adult cardiomyocyte. Proc. Natl Acad. Sci. USA 95, 2979–2984 (1998).

    Article  CAS  Google Scholar 

  22. Shuralyova, I. et al. Inhibition of glucose uptake in murine cardiomyocyte cell line HL-1 by cardioprotective drugs dilazep and dipyridamole. Am. J. Physiol. Heart Circ. Physiol. 286, H627–H632 (2004).

    Article  CAS  Google Scholar 

  23. Hay, J. C. et al. Localization, dynamics, and protein interactions reveal distinct roles for ER and Golgi SNAREs. J. Cell Biol. 141, 1489–1502 (1998).

    Article  CAS  Google Scholar 

  24. Lanoix, J. et al. Sorting of Golgi resident proteins into different subpopulations of COPI vesicles: a role for ArfGAP1. J. Cell Biol. 155, 1199–1212 (2001).

    Article  CAS  Google Scholar 

  25. Sollner, T. et al. A protein assembly-disassembly pathway in vitro that may correspond to sequential steps of synaptic vesicle docking, activation, and fusion. Cell 75, 409–418 (1993).

    Article  CAS  Google Scholar 

  26. Asp, L. et al. Role of ADP ribosylation factor 1 in the assembly and secretion of ApoB-100-containing lipoproteins. Arterioscler. Thromb. Vasc. Biol. 25, 566–570 (2005).

    Article  CAS  Google Scholar 

  27. Piper, R. C., Hess, L. J., & James, D. E. Differential sorting of two glucose transporters expressed in insulin-sensitive cells. Am. J. Physiol. 260, C570–C580 (1991).

    Article  CAS  Google Scholar 

  28. Olefsky, J. M. Mechanisms of the ability of insulin to activate the glucose-transport system in rat adipocytes. Biochem. J. 172, 137–145 (1978).

    Article  CAS  Google Scholar 

Download references

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.

Author information

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

Authors
  1. Pontus Boström
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Linda Andersson
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mikael Rutberg
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jeanna Perman
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ulf Lidberg
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Bengt R. Johansson
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Julia Fernandez-Rodriguez
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Johanna Ericson
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Tommy Nilsson
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Jan Borén
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Sven-Olof Olofsson
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Sven-Olof Olofsson.

Supplementary information

Supplementary Information

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

Rights and permissions

Reprints and permissions

About this article

Cite this article

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

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ncb1648

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing