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.

  • Brief Communication
  • Published:

Activation of membrane-permeant caged PtdIns(3)P induces endosomal fusion in cells

Nature Chemical Biology volume 6, pages 324–326 (2010)Cite this article

Subjects

Abstract

Phosphatidylinositol 3-phosphate (PtdIns(3)P) is a phospholipid residing on early endosomes, where it is proposed to be involved in endosomal fusion. We synthesized membrane-permeant derivatives of PtdIns(3)P, including a caged version that is to our knowledge the first photoactivatable phosphoinositide derivative developed so far. In living cells, photoactivation of caged PtdIns(3)P induced rapid endosomal fusion in an EEA1-dependent fashion, thus providing in vivo evidence that PtdIns(3)P is a sufficient signal for driving this process.

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: PtdIns(3)P/AM-induced endosome fusion.

Similar content being viewed by others

References

  1. Cantley, L.C. Science 296, 1655–1657 (2002).

    Article  CAS  Google Scholar 

  2. Simonsen, A., Wurmser, A.E., Emr, S.D. & Stenmark, H. Curr. Opin. Cell Biol. 13, 485–492 (2001).

    Article  CAS  Google Scholar 

  3. Roth, M.G. Physiol. Rev. 84, 699–730 (2004).

    Article  CAS  Google Scholar 

  4. Schu, P.V. et al. Science 260, 88–91 (1993).

    Article  CAS  Google Scholar 

  5. Gillooly, D.J. et al. EMBO J. 19, 4577–4588 (2000).

    Article  CAS  Google Scholar 

  6. Jones, A.T. & Clague, M.J. Biochem. J. 311, 31–34 (1995).

    Article  CAS  Google Scholar 

  7. Wurmser, A.E., Gary, J.D. & Emr, S.D. J. Biol. Chem. 274, 9129–9132 (1999).

    Article  CAS  Google Scholar 

  8. Vergne, I., Chua, J. & Deretic, V. Traffic 4, 600–606 (2003).

    Article  CAS  Google Scholar 

  9. Fratti, R.A., Backer, J.M., Gruenberg, J., Corvera, S. & Deretic, V. J. Cell Biol. 154, 631–644 (2001).

    Article  CAS  Google Scholar 

  10. Petiot, A., Ogier-Denis, E., Blommaart, E.F., Meijer, A.J. & Codogno, P. J. Biol. Chem. 275, 992–998 (2000).

    Article  CAS  Google Scholar 

  11. Blommaart, E.F., Krause, U., Schellens, J.P., Vreeling-Sindelarova, H. & Meijer, A.J. Eur. J. Biochem. 243, 240–246 (1997).

    Article  CAS  Google Scholar 

  12. Xie, Z. & Klionsky, D.J. Nat. Cell Biol. 9, 1102–1109 (2007).

    Article  CAS  Google Scholar 

  13. Dinkel, C., Moody, M., Traynor-Kaplan, A. & Schultz, C. Angew. Chem. Int. Edn Engl. 40, 3004–3008 (2001).

    Article  CAS  Google Scholar 

  14. Jiang, T. et al. J. Biol. Chem. 273, 11017–11024 (1998).

    Article  CAS  Google Scholar 

  15. Laketa, V. et al. Chem. Biol. 16, 1190–1196 (2009).

    Article  CAS  Google Scholar 

  16. Walker, J.W., Reid, G.P., McCray, J.A. & Trentham, D.R. J. Am. Chem. Soc. 110, 7170–7177 (1988).

    Article  CAS  Google Scholar 

  17. Hagen, V. et al. Angew. Chem. Int. Edn Engl. 40, 1045–1048 (2001).

    Article  Google Scholar 

  18. Walker, J.W., Somlyo, A.V., Goldman, Y.E., Somlyo, A.P. & Trentham, D.R. Nature 327, 249–252 (1987).

    Article  CAS  Google Scholar 

  19. Dinkel, C. & Schultz, C. Tetrahedr. Lett. 44, 1157–1159 (2003).

    Article  CAS  Google Scholar 

  20. Li, W., Llopis, J., Whitney, M., Zlokarnik, G. & Tsien, R.Y. Nature 392, 936–941 (1998).

    Article  CAS  Google Scholar 

  21. Kantevari, S. et al. Chembiochem. 7, 174–180 (2006).

    Article  CAS  Google Scholar 

  22. Li, G. et al. Proc. Natl. Acad. Sci. USA 92, 10207–10211 (1995).

    Article  CAS  Google Scholar 

  23. Ohya, T. et al. Nature 459, 1091–1097 (2009).

    Article  CAS  Google Scholar 

  24. Simonsen, A. et al. Nature 394, 494–498 (1998).

    Article  CAS  Google Scholar 

  25. Lawe, D.C. et al. J. Biol. Chem. 277, 8611–8617 (2002).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank the Advanced Light Microscopy Facility of the European Molecular Biology Laboratory (EMBL), and we also thank C. Funaya and C. Antony of the EMBL Electron Microscopy Core Facility for EM sample preparation. Funding was provided by the Volkswagen Foundation (I81/597 and I81/797) and the Helmholtz-Initiative on Systems Biology (SBCancer) to C.S.

Author information

Author notes

  1. Devaraj Subramanian and Vibor Laketa: These authors contributed equally to this work.

Authors and Affiliations

  1. European Molecular Biology Institute, Cell Biology and Biophysics Unit, Heidelberg, Germany

    Devaraj Subramanian, Vibor Laketa, Rainer Müller, Christian Tischer, Sirus Zarbakhsh, Rainer Pepperkok & Carsten Schultz

Authors
  1. Devaraj Subramanian
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Vibor Laketa
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Rainer Müller
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Christian Tischer
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Sirus Zarbakhsh
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Rainer Pepperkok
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Carsten Schultz
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

D.S., V.L., S.Z., R.P. and C.S. designed the experiments and wrote the manuscript. D.S., V.L., R.M. and S.Z. performed the experiments and analyzed the data. C.T. developed the analytical software.

Corresponding author

Correspondence to Carsten Schultz.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–7 and Supplementary Methods (PDF 2502 kb)

Supplementary Video 1

PtdIns(3)P/AM induced endosome fusion. HeLa cells were analyzed by time-lapse microscopy as described in Supplementary Methods. They were treated with PtdIns(3)P/AM (100 μM) at time point 3min. Image was acquired every 30 seconds during 25 minutes. Note the appearance of large endosomes. Time is indicated in the upper left corner. Scale bar, 10 μm. (AVI 4167 kb)

Supplementary Video 2

caged-PtdIns(3)P/AM induced endosome fusion. HeLa cells were analyzed by time-lapse microscopy as described in Supplementary Methods. They were treated with caged-PtdIns(3)P/AM (100 μM) for 10 min before the UV flash (time point 0min). Image was acquired every 30 seconds during 25 minutes. Note the appearance of large endosomes. Time is indicated in the upper left corner. Scale bar, 10 μm. (AVI 4065 kb)

Supplementary Video 3

PtdIns(4)P/AM did not induce endosome fusion. HeLa cells were analyzed by time-lapse microscopy as described in Supplementary Methods. They were treated with PtdIns(4)P/AM (100 μM) at time point 4min. Image was acquired every 30 seconds during 50 minutes. Time is indicated in the upper left corner. Scale bar, 10 μm. (AVI 5111 kb)

Supplementary Video 4

UV flash did not induce endosome fusion. HeLa cells were analyzed by time-lapse microscopy as described in Supplementary Methods. They were illuminated with UV light at time point 0 min. Image was acquired every 30 seconds during 35 minutes. Time is indicated in the upper left corner. Scale bar, 10 μm. (AVI 5030 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Subramanian, D., Laketa, V., Müller, R. et al. Activation of membrane-permeant caged PtdIns(3)P induces endosomal fusion in cells. Nat Chem Biol 6, 324–326 (2010). https://doi.org/10.1038/nchembio.348

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nchembio.348

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