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:

COPII–cargo interactions direct protein sorting into ER-derived transport vesicles

Nature volume 391, pages 187–190 (1998)Cite this article

Abstract

Vesicles coated with coat protein complex II (COPII) selectively transport molecules (cargo) and vesicle fusion proteins from the endoplasmic reticulum (ER) to the Golgi complex1,2,3. We have investigated the role of coat proteins in cargo selection and recruitment. We isolated integral membrane and soluble cargo proteins destined for transport from the ER in complexes formed in the presence of Sar1 and Sec23/24, a subset of the COPII components, and GTP or GMP-PNP. Vesicle fusion proteins of the vSNARE family and Emp24, a member of a putative cargo carrier family4, were also found in COPII complexes. The inclusion of amino-acid permease molecules into the complex depended on the presence of Shr3, a protein required for the permease to leave the ER3,5. Resident ER proteins Sec61, BiP (Kar2) and Shr3 were not included in the complexes, indicating that the COPII components bound specifically to vesicle cargo. COPII–cargo complexes and putative cargo adaptor–cargo complexes were also isolated from COPII vesicles. Our results indicate that cargo packaging signals and soluble cargo adaptors are recognized by a recruitment complex comprising Sar1–GTP and Sec23/24.

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: Vesicle proteins Hip1, Emp24, Sec22 and gpαF, but not ER resident proteins Sec61, Shr3 and Kar2, are present in complex with Sar1 and Sec23/24.
Figure 2: Vesicle proteins Hip1, Emp24, Sec22 and gpαF, but not ER resident proteins Sec61, Shr3 and Kar2, are present in complex with Sar1 and Sec23/24.
Figure 3: Shr3 is required specifically for incorporation of Hip1 and Gap1 into a complex with Sar1 and Sec23/24.
Figure 4: Shr3 is required specifically for incorporation of Hip1 and Gap1 into a complex with Sar1 and Sec23/24.
Figure 5: Crosslinking of gpαF in a prebudding complex from ER membranes and ER-derived transport vesicles.
Figure 6: Crosslinking of gpαF in a prebudding complex from ER membranes and ER-derived transport vesicles.
Figure 7: Crosslinking of gpαF in a prebudding complex from ER membranes and ER-derived transport vesicles.

Similar content being viewed by others

References

  1. Salama, N. R., Yeung, T. & Schekman, R. W. The Sec13p complex and reconstitution of vesicle budding from the ER with purified cytosolic proteins. EMBO J. 12, 4073–4082 (1993).

    Article  CAS  Google Scholar 

  2. Barlow, 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  Google Scholar 

  3. Kuehn, M. J., Schekman, R. & Ljungdahl, P. O. Amino acid permeases require COPII components and the ER resident membrane protein Shr3p for packaging into transport vesicles in vitro. J. Cell Biol. 135, 585–595 (1996).

    Article  CAS  Google Scholar 

  4. Schimmöller, F. et al. The absence of Emp24p, a component of ER-derived COPII-coated vesicles, causes a defect in transport of selected proteins to the Golgi. EMBO J. 14, 1329–1339 (1995).

    Article  Google Scholar 

  5. Ljungdahl, P. O., Gimeno, C. J., Styles, C. A. & Fink, G. R. SHR3: a novel component of the secretory pathway specifically required for localization of amino acid permeases in yeast. Cell 71, 463–478 (1992).

    Article  CAS  Google Scholar 

  6. Barlowe, C. & Schekman, R. SEC12 encodes a guanine-nucleotide-exchange factor essential for transport vesicle budding from the ER. Nature 365, 347–349 (1993).

    Article  ADS  CAS  Google Scholar 

  7. 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 

  8. Yeung, T., Barlowe, C. & Schekman, R. Uncoupled packaging of targeting and cargo molecules during transport vesicle budding from the endoplasmic reticulum. J. Biol. Chem. 270, 30567–30570 (1995).

    Article  CAS  Google Scholar 

  9. Kuehn, M. J. & Schekman, R. COPII and secretory cargo capture into transport vesicles. Curr. Opin. Cell Biol. 9, 477–483 (1997).

    Article  CAS  Google Scholar 

  10. Stamnes, J. A. et al. An integral membrane component of coatomer-coated transport vesicles defines a family of proteins involved in budding. Proc. Natl Acad. Sci. USA 92, 8011–8015 (1995).

    Article  ADS  CAS  Google Scholar 

  11. Fiedler, K., Veit, M., Stamnes, M. A. & Rothman, J. E. Bimodal interaction of coatomer with the p24 family of putative cargo receptors. Science 273, 1396–1399 (1996).

    Article  ADS  CAS  Google Scholar 

  12. Barlow, C., d'Enfert, C. & Schekman, R. Purification and characterization of Sar1p, a small GTP binding protein required for transport vesicle formation from the endoplasmic reticulum. J. Biol. Chem. 268, 873–879 (1993).

    Google Scholar 

  13. Fiedler, K. & Simons, K. The role of N-glycans in the secretory pathway. Cell 81, 309–312 (1995).

    Article  CAS  Google Scholar 

  14. Sikorski, R. S. & Heiter, P. Asystem of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics 122, 19–27 (1989).

    CAS  PubMed  PubMed Central  Google Scholar 

  15. Hicke, L., Yoshihisa, T. & Schekman, R. Sec23p and a novel 105 kD protein function as a multimeric complex to promote vesicle budding from the ER. Mol. Biol. Cell 3, 667–676 (1992).

    Article  CAS  Google Scholar 

  16. Stirling, C. J., Rothblatt, J., Hosobuchi, M., Deshaies, R. & Schekman, R. Protein translocation mutants defective in the insertion of integral membrane proteins into the endoplasmic reticulum. Mol. Biol. Cell 3, 129–142 (1992).

    Article  CAS  Google Scholar 

  17. Brodsky, J. L. & Schekman, R. ASec63p-BIP complex from yeast is required for protein translocation in a reconstituted proteoliposome. J. Cell Biol. 123, 1355–1363 (1993).

    Article  CAS  Google Scholar 

  18. Bednarek, S. Y. et al. COPI- and COPII-coated vesicles bud directly from the endoplasmic reticulum in yeast. Cell 83, 1183–1196 (1995).

    Article  CAS  Google Scholar 

  19. Evan, G. I., Lewis, G. K., Ramsay, G. & Bishop, J. M. Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product. Mol. Cell. Biol. 5, 3610–3616 (1985).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank S. Bednarek, M. Bowser and L. Liang for performing experiments; P.Ljungdahl, R. Lesch, M. Pilon and A. Corsi for materials; and J. Campbell, T. Doering and other lab members for suggestions. This work was supported by the Howard Hughes Medical Institute (R.S. and M.K.) and the Cancer Research Fund fo the Damon Runyon-Walter Winchell Foundation (M.K.). J.H. was supported by a fellowship from the Deutsche Forschungsgemeinschaft.

Author information

Authors and Affiliations

  1. Department of Molecular and Cell Biology, Barker Hall, Howard Hughes Medical Institute, University of California, Berkeley, California, 94720, USA

    Meta J. Kuehn, Johannes M. Herrmann & Randy Schekman

Authors
  1. Meta J. Kuehn
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Johannes M. Herrmann
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Randy Schekman
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Randy Schekman.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kuehn, M., Herrmann, J. & Schekman, R. COPII–cargo interactions direct protein sorting into ER-derived transport vesicles. Nature 391, 187–190 (1998). https://doi.org/10.1038/34438

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/34438

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