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.

  • Article
  • Published:

Sequential interactions with Sec23 control the direction of vesicle traffic

Nature volume 473, pages 181–186 (2011)Cite this article

Subjects

Abstract

How the directionality of vesicle traffic is achieved remains an important unanswered question in cell biology. The Sec23p/Sec24p coat complex sorts the fusion machinery (SNAREs) into vesicles as they bud from the endoplasmic reticulum (ER). Vesicle tethering to the Golgi begins when the tethering factor TRAPPI binds to Sec23p. Where the coat is released and how this event relates to membrane fusion is unknown. Here we use a yeast transport assay to demonstrate that an ER-derived vesicle retains its coat until it reaches the Golgi. A Golgi-associated kinase, Hrr25p (CK1δ orthologue), then phosphorylates the Sec23p/Sec24p complex. Coat phosphorylation and dephosphorylation are needed for vesicle fusion and budding, respectively. Additionally, we show that Sec23p interacts in a sequential manner with different binding partners, including TRAPPI and Hrr25p, to ensure the directionality of ER–Golgi traffic and prevent the back-fusion of a COPII vesicle with the ER. These events are conserved in mammalian cells.

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: COPII vesicles lose their ability to bind TRAPPI after Uso1p acts.
Figure 2: Hrr25p resides on the Golgi and phosphorylates Sec23p/Sec24p.
Figure 3: Phosphorylation of S742 and T747 blocks ER–Golgi traffic in vitro.
Figure 4: TRAPPI and Sar1p–GTP cannot bind to Sec23p simultaneously.
Figure 5: In the presence of IC261, cargo is exported more rapidly but remains at peripheral pre-Golgi sites.
Figure 6: Sec23p ensures the direction of ER–Golgi traffic.

Similar content being viewed by others

References

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

    Article  CAS  Google Scholar 

  2. Lewis, M. J., Rayner, J. C. & Pelham, H. R. A novel SNARE complex implicated in vesicle fusion with the endoplasmic reticulum. EMBO J. 16, 3017–3024 (1997)

    Article  CAS  Google Scholar 

  3. Mossessova, E., Bickford, L. C. & Goldberg, J. SNARE selectivity of the COPII coat. Cell 114, 483–495 (2003)

    Article  CAS  Google Scholar 

  4. Lee, M. C. S., Miller, E. A., Goldberg, J., Orci, L. & Schekman, R. Bi-directional protein transport between the ER and Golgi. Annu. Rev. Cell Dev. Biol. 20, 87–123 (2004)

    Article  CAS  Google Scholar 

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

    CAS  PubMed  Google Scholar 

  6. Cai, H. et al. TRAPPI tethers COPII vesicles by binding the coat subunit Sec23. Nature 445, 941–944 (2007)

    Article  CAS  ADS  Google Scholar 

  7. 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 

  8. Flanagan, J. J. & Barlowe, C. Cysteine-disulfide cross-linking to monitor SNARE complex assembly during endoplasmic reticulum-Golgi transport. J. Biol. Chem. 281, 2281–2288 (2006)

    Article  CAS  Google Scholar 

  9. Dudognon, P., Maeder-Garavaglia, C., Carpentier, J. & Paccaud, J. Regulation of a COPII component by cytosolic O-glycosylation during mitosis. FEBS Lett. 561, 44–50 (2004)

    Article  CAS  Google Scholar 

  10. Milne, D. M., Looby, P. & Meek, D. W. Catalytic activity of protein kinase CK1δ (casein kinase 1δ) is essential for its normal subcellular localization. Exp. Cell Res. 263, 43–54 (2001)

    Article  CAS  Google Scholar 

  11. Yu, S. & Roth, M. G. Casein kinase I regulates membrane binding by ARF GAP1. Mol. Biol. Cell 13, 2559–2570 (2002)

    Article  CAS  Google Scholar 

  12. Murakami, A., Kimura, K. & Nakano, A. The inactive form of a yeast casein kinase I suppresses the secretory defect of the sec12 mutant. J. Biol. Chem. 274, 3804–3810 (1999)

    Article  CAS  Google Scholar 

  13. Lusk, C. P. et al. Nup53p is a target of two mitotic kinases, Cdk1p and Hrr25p. Traffic 8, 647–660 (2007)

    Article  CAS  Google Scholar 

  14. Kafadar, K. A., Zhu, H., Snyder, M. & Cyert, M. S. Negative regulation of calcineurin signaling by Hrr25p, a yeast homolog of casein kinase I. Genes Dev. 17, 2698–2708 (2003)

    Article  CAS  Google Scholar 

  15. Bi, X., Corpina, R. A. & Goldberg, J. Structure of the Sec23/24-Sar1 pre-budding complex of the COPII vesicle coat. Nature 419, 271–277 (2002)

    Article  CAS  ADS  Google Scholar 

  16. Mashhoon, N. et al. Crystal structure of a conformation-selective casein kinase-1 inhibitor. J. Biol. Chem. 275, 20052–20060 (2000)

    Article  CAS  Google Scholar 

  17. 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 

  18. Schweizer, A. et al. Identification of an intermediate compartment involved in protein transport from endoplasmic reticulum to Golgi apparatus. Eur. J. Cell Biol. 53, 185–196 (1990)

    CAS  PubMed  Google Scholar 

  19. Bielli, A. et al. Regulation of Sar1 NH2 terminus by GTP binding and hydrolysis promotes membrane deformation to control COPII vesicle fission. J. Cell Biol. 171, 919–924 (2005)

    Article  CAS  Google Scholar 

  20. Lee, M. C. S. et al. Sar1p N-terminal helix initiates membrane curvature and completes the fission of a COPII vesicle. Cell 122, 605–617 (2005)

    Article  CAS  Google Scholar 

  21. Chin, H. F. et al. Kinetic analysis of the guanine nucleotide exchange activity of TRAPP, a multimeric Ypt1p exchange factor. J. Mol. Biol. 389, 275–288 (2009)

    Article  CAS  Google Scholar 

  22. Salama, N. R., Chuang, J. S. & Schekman, R. W. Sec31 encodes an essential component of the COPII coat required for transport vesicle budding from the endoplasmic reticulum. Mol. Biol. Cell 8, 205–217 (1997)

    Article  CAS  Google Scholar 

  23. Guttman, M. et al. Interactions of the NPXY microdomains of the low density lipoprotein receptor-related protein 1. Proteomics 9, 5016–5028 (2009)

    Article  CAS  Google Scholar 

  24. Gimeno, R. E., Espenshade, P. & Kaiser, C. A. COPII coat subunit interactions: Sec24p and Sec23p bind to adjacent regions of Sec16p. Mol. Biol. Cell 7, 1815–1823 (1996)

    Article  CAS  Google Scholar 

  25. Fromme, J. C. et al. The genetic basis of a craniofacial disease provides insight into COPII coat assembly. Dev. Cell 13, 623–634 (2007)

    Article  CAS  Google Scholar 

  26. Diao, A., Frost, L., Morohashi, Y. & Lowe, M. Coordination of golgin tethering and SNARE assembly: GM130 binds syntaxin 5 in a p115-regulated manner. J. Biol. Chem. 283, 6957–6967 (2008)

    Article  CAS  Google Scholar 

  27. Saraste, J. & Svensson, K. Distribution of the intermediate elements operating in ER to Golgi transport. J. Cell Sci. 100, 415–430 (1991)

    CAS  PubMed  Google Scholar 

  28. 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 

  29. Bentley, M. et al. Vesicular calcium regulates coat retention, fusogenicity, and size of pre-Golgi intermediates. Mol. Biol. Cell 21, 1033–1046 (2010)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank R. Schekman, M. Cyert, L. Miller, B. Glick, M. Lowe and E. Mizuno-Yamasaki for strains, constructs, purified protein and antibody; K. Reinisch for discussions; S. Chen, H. Cai and M. Garcia-Marcos for advice; W. Zhou and A. Lougheed for technical assistance. This work was supported by the Howard Hughes Medical Institute. Salary support for S.F.-N., D.B. and S.M. was provided by the Howard Hughes Medical Institute, P.G. was funded by the Burroughs Wellcome Fund and M.G. by the SRP Super Fund Research Program. Work at the University of Montana was supported by National Institutes of Health grant GM-059378 (to J.C.H.) and the National Institutes of Health COBRE Center grant RR-015583.

Author information

Author notes

  1. Christopher Lord and Deepali Bhandari: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Cellular and Molecular Medicine, Howard Hughes Medical Institute, University of California at San Diego, La Jolla, 92093-0668, California, USA

    Christopher Lord, Deepali Bhandari, Shekar Menon & Susan Ferro-Novick

  2. Department of Chemistry and Biochemistry, Biomolecular and Proteomics Mass Spectrometry Facility, University of California at San Diego, La Jolla, 92093, California, USA

    Majid Ghassemian

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

    Deborah Nycz & Jesse Hay

  4. Department of Medicine, University of California at San Diego, La Jolla, 92093-0651, California, USA

    Pradipta Ghosh

Authors
  1. Christopher Lord
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Deepali Bhandari
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Shekar Menon
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Majid Ghassemian
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Deborah Nycz
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jesse Hay
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Pradipta Ghosh
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Susan Ferro-Novick
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

C.L., D.B., S.M., D.N. and J.H. performed experiments and analysed data. M.G. did the mass analysis and P.G. did the computational modelling and analysed data. S.F.-N. directed the project, analysed data and wrote the paper. C.L., D.B., J.H. and P.G. co-wrote the paper.

Corresponding author

Correspondence to Susan Ferro-Novick.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

The file contains Supplementary Figures 1-5 with legends and an additional reference. (PDF 596 kb)

PowerPoint slides

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lord, C., Bhandari, D., Menon, S. et al. Sequential interactions with Sec23 control the direction of vesicle traffic. Nature 473, 181–186 (2011). https://doi.org/10.1038/nature09969

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature09969

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

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