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TRAPPI tethers COPII vesicles by binding the coat subunit Sec23

Nature volume 445, pages 941–944 (2007)Cite this article

Abstract

The budding of endoplasmic reticulum (ER)-derived vesicles is dependent on the COPII coat complex1. Coat assembly is initiated when Sar1-GTP recruits the cargo adaptor complex, Sec23/Sec24, by binding to its GTPase-activating protein (GAP) Sec23 (ref. 2). This leads to the capture of transmembrane cargo by Sec24 (refs 3, 4) before the coat is polymerized by the Sec13/Sec31 complex5. The initial interaction of a vesicle with its target membrane is mediated by tethers6. We report here that in yeast and mammalian cells the tethering complex TRAPPI (ref. 7) binds to the coat subunit Sec23. This event requires the Bet3 subunit. In vitro studies demonstrate that the interaction between Sec23 and Bet3 targets TRAPPI to COPII vesicles to mediate vesicle tethering. We propose that the binding of TRAPPI to Sec23 marks a coated vesicle for fusion with another COPII vesicle or the Golgi apparatus. An implication of these findings is that the intracellular destination of a transport vesicle may be determined in part by its coat and its associated cargo.

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Figure 1: mBet3 tethers coated vesicles in mammalian cells.
Figure 2: The interaction between Sec23p and Bet3p is required for the binding of COPII vesicles to yTRAPPI.
Figure 3: Sec23p is required for COPII vesicle tethering.
Figure 4: yTRAPPI links vesicle recognition to Rab activation.

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References

  1. Barlowe, C. et al. COPII: a membrane coat formed by Sec proteins that drive vesicle budding from the endoplasmic reticulum. Cell 77, 895–907 (1994)

    Article  CAS  Google Scholar 

  2. Yoshihisa, T., Barlowe, C. & Schekman, R. Requirement for a GTPase-activating protein in vesicle budding from the endoplasmic reticulum. Science 259, 1466–1468 (1993)

    Article  ADS  CAS  Google Scholar 

  3. Kuehn, M. J., Herrmann, J. M. & Schekman, R. COPII-cargo interactions direct protein sorting into ER-derived transport vesicles. Nature 391, 187–190 (1998)

    Article  ADS  CAS  Google Scholar 

  4. Miller, E. A. et al. Multiple cargo binding sites on the COPII subunit Sec24p ensure capture of diverse membrane proteins into transport vesicles. Cell 114, 497–509 (2003)

    Article  CAS  Google Scholar 

  5. Stagg, S. M. et al. Structure of the Sec13/31 COPII coat cage. Nature 439, 234–238 (2006)

    Article  ADS  CAS  Google Scholar 

  6. Whyte, J. R. & Munro, S. Vesicle tethering complexes in membrane traffic. J. Cell Sci. 115, 2627–2637 (2002)

    CAS  PubMed  Google Scholar 

  7. Sacher, M. et al. TRAPP I implicated in the specificity of tethering in ER-to-Golgi transport. Mol. Cell 7, 433–442 (2001)

    Article  CAS  Google Scholar 

  8. Cao, X., Ballew, N. & Barlowe, C. Initial docking of ER-derived vesicles requires Uso1p and Ypt1p but is independent of SNARE proteins. EMBO J. 17, 2156–2165 (1998)

    Article  CAS  Google Scholar 

  9. Sacher, M. et al. TRAPP, a highly conserved novel complex on the cis-Golgi that mediates vesicle docking and fusion. EMBO J. 17, 2494–2503 (1998)

    Article  CAS  Google Scholar 

  10. Yu, S. et al. mBet3p is required for homotypic COPII vesicle tethering in mammalian cells. J. Cell Biol. 174, 359–368 (2006)

    Article  CAS  Google Scholar 

  11. Quintyne, N. J. et al. Dynactin is required for microtubule anchoring at centrosomes. J. Cell Biol. 147, 321–334 (1999)

    Article  CAS  Google Scholar 

  12. Xu, D. & Hay, J. C. Reconstitution of COPII vesicle fusion to generate a pre-Golgi intermediate compartment. J. Cell Biol. 167, 997–1003 (2004)

    Article  CAS  Google Scholar 

  13. Peyroche, A. & Jackson, C. L. Functional analysis of ADP-ribosylation factor (ARF) guanine nucleotide exchange factors Gea1p and Gea2p in yeast. Methods Enzymol. 329, 290–300 (2001)

    Article  CAS  Google Scholar 

  14. Antonny, B., Madden, D., Hamamoto, S., Orci, L. & Schekman, R. Dynamics of the COPII coat with GTP and stable analogues. Nature Cell Biol. 3, 531–537 (2001)

    Article  CAS  Google Scholar 

  15. Barrowman, J., Sacher, M. & Ferro-Novick, S. TRAPP stably associates with the Golgi and is required for vesicle docking. EMBO J. 19, 862–869 (2000)

    Article  CAS  Google Scholar 

  16. Rexach, M. F. & Schekman, R. W. Distinct biochemical requirements for the budding, targeting, and fusion of ER-derived transport vesicles. J. Cell Biol. 114, 219–229 (1991)

    Article  CAS  Google Scholar 

  17. Oka, T. & Nakano, A. Inhibition of GTP hydrolysis by Sar1p causes accumulation of vesicles that are a functional intermediate of the ER-to-Golgi transport in yeast. J. Cell Biol. 124, 425–434 (1994)

    Article  CAS  Google Scholar 

  18. Sato, K. & Nakano, A. Dissection of COPII subunit-cargo assembly and disassembly kinetics during Sar1p-GTP hydrolysis. Nature Struct. Mol. Biol. 12, 167–174 (2005)

    Article  CAS  Google Scholar 

  19. Forster, R. et al. Secretory cargo regulates the turnover of COPII subunits at single ER exit sites. Curr. Biol. 16, 173–179 (2006)

    Article  CAS  Google Scholar 

  20. Menon, S. et al. mBet3 is required for the organization of the TRAPP complexes. Biochem. Biophys. Res. Commun. 350, 669–677 (2006)

    Article  CAS  Google Scholar 

  21. Wang, W., Sacher, M. & Ferro-Novick, S. TRAPP stimulates guanine nucleotide exchange on Ypt1p. J. Cell Biol. 151, 289–296 (2000)

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  23. Söllner, T. et al. SNAP receptors implicated in vesicle targeting and fusion. Nature 362, 318–324 (1993)

    Article  ADS  Google Scholar 

  24. Rothman, J. E. Mechanisms of intracellular protein transport. Nature 372, 55–63 (1994)

    Article  ADS  CAS  Google Scholar 

  25. Shorter, J., Beard, M. B., Seemann, J., Dirac-Svejstrup, A. B. & Warren, G. Sequential tethering of Golgins and catalysis of SNAREpin assembly by the vesicle-tethering protein p115. J. Cell Biol. 157, 45–62 (2002)

    Article  CAS  Google Scholar 

  26. Bentley, M., Liang, Y., Mullen, K., Xu, D., Sztul, E. & Hay, J. C. SNARE status regulates tether recruitment and function in homotypic COPII vesicle fusion. J. Biol. Chem. 281, 38825–38833 (2006)

    Article  CAS  Google Scholar 

  27. Kim, Y.-G. et al. The architecture of the multisubunit TRAPP I complex suggests a model for vesicle tethering. Cell 127, 817–830 (2006)

    Article  CAS  Google Scholar 

  28. Sacher, M., Barrowman, J., Schieltz, D., Yates, J. R. & Ferro-Novick, S. Identification and characterization of five new subunits of TRAPP. Eur. J. Cell Biol. 79, 71–80 (2000)

    Article  CAS  Google Scholar 

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Acknowledgements

We thank L. Walker for help in the preparation of the manuscript, Y. Zhang, and K. Mullen for technical assistance. We also thank J. Barrowman for help with the yeast in vitro tethering assay, R. Schekman, C. Kaiser, F. Gorelick and T. Schroer for purified proteins, antibodies, plasmids and strains, and E. Miller for advice on stripping COPII coats from vesicles. This work was supported by the Howard Hughes Medical Institute. Salary support for S.F.-N., H.C., S.Y., S.M. and D.L. was also provided by the Howard Hughes Medical Institute. Work at the University of Montana was supported by an NIH grant to J.C.H. and the NIH-COBRE Center for Structural and Functional Neuroscience.

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  1. Howard Hughes Medical Institute and the,,

    Huaqing Cai, Sidney Yu, Shekar Menon, Darina Lazarova & Susan Ferro-Novick

  2. Department of Cell Biology Yale University School of Medicine, New Haven, Connecticut 06519, USA,

    Huaqing Cai, Sidney Yu, Shekar Menon, Yiying Cai, Chunmei Fu, Karin Reinisch & Susan Ferro-Novick

  3. Division of Biological Sciences, The University of Montana, Missoula, Montana 59812-4824, USA,

    Jesse C. Hay

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Correspondence to Susan Ferro-Novick.

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Cai, H., Yu, S., Menon, S. et al. TRAPPI tethers COPII vesicles by binding the coat subunit Sec23. Nature 445, 941–944 (2007). https://doi.org/10.1038/nature05527

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