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Molecular organization of the COG vesicle tethering complex

Nature Structural & Molecular Biology volume 17, pages 1292–1297 (2010)Cite this article

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Abstract

Multisubunit tethering complexes of the CATCHR (complexes associated with tethering containing helical rods) family are proposed to mediate the initial contact between an intracellular trafficking vesicle and its membrane target. To begin elucidating the molecular architecture of one well-studied example, the conserved oligomeric Golgi (COG) complex, we reconstituted its essential subunits (Cog1, Cog2, Cog3 and Cog4) and used single-particle electron microscopy to reveal a y-shaped structure with three flexible, highly extended legs. Labeling experiments established that the N termini of all four subunits interact along the proximal segment of one leg, whereas three of the four C termini are located at the tips of the legs. Our results suggest that the central region of the Cog1–Cog2–Cog3–Cog4 complex, as well as the distal regions of at least two legs, all participate in interactions with other components of the intracellular trafficking machinery.

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Figure 1: Analysis of the quadruple knockout strain for growth and trafficking defects.
Figure 2: Purification and electron microscopy of the COG subcomplexes.
Figure 3: Projection structures of the Cog2–4 and Cog1–4 (core) complexes.
Figure 4: Model for subunit organization in the Cog1–4 core complex.
Figure 5: Functional architecture of the Cog1–4 core complex.

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References

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

    CAS  Google Scholar 

  2. Cai, H., Reinisch, K. & Ferro-Novick, S. Coats, tethers, Rabs, and SNAREs work together to mediate the intracellular destination of a transport vesicle. Dev. Cell 12, 671–682 (2007).

    Article  CAS  Google Scholar 

  3. Sztul, E. & Lupashin, V. Role of vesicle tethering factors in the ER-Golgi membrane traffic. FEBS Lett. 583, 3770–3783 (2009).

    Article  CAS  Google Scholar 

  4. Yu, I. & Hughson, F.M. Tethering factors as organizers of intracellular vesicular traffic. Annu. Rev. Cell Dev. Biol. 26, 137–156 (2010).

    Article  CAS  Google Scholar 

  5. Ungar, D., Oka, T., Krieger, M. & Hughson, F.M. Retrograde transport on the COG railway. Trends Cell Biol. 16, 113–120 (2006).

    Article  CAS  Google Scholar 

  6. Ungar, D. et al. Characterization of a mammalian Golgi-localized protein complex, COG, that is required for normal Golgi morphology and function. J. Cell Biol. 157, 405–415 (2002).

    Article  CAS  Google Scholar 

  7. Walter, D.M., Paul, K.S. & Waters, M.G. Purification and characterization of a novel 13 S hetero–oligomeric protein complex that stimulates in vitro Golgi transport. J. Biol. Chem. 273, 29565–29576 (1998).

    Article  CAS  Google Scholar 

  8. Whyte, J.R. & Munro, S. The Sec34/35 Golgi transport complex is related to the exocyst, defining a family of complexes involved in multiple steps of membrane traffic. Dev. Cell 1, 527–537 (2001).

    Article  CAS  Google Scholar 

  9. VanRheenen, S.M., Cao, X., Lupashin, V.V., Barlowe, C. & Waters, M.G. Sec35p, a novel peripheral membrane protein, is required for ER to Golgi vesicle docking. J. Cell Biol. 141, 1107–1119 (1998).

    Article  CAS  Google Scholar 

  10. VanRheenen, S.M. et al. Sec34p, a protein required for vesicle tethering to the yeast Golgi apparatus, is in a complex with Sec35p. J. Cell Biol. 147, 729–742 (1999).

    Article  CAS  Google Scholar 

  11. Wuestehube, L.J. et al. New mutants of Saccharomyces cerevisiae affected in the transport of proteins from the endoplasmic reticulum to the Golgi complex. Genetics 142, 393–406 (1996).

    CAS  PubMed  PubMed Central  Google Scholar 

  12. Ram, R.J., Li, B. & Kaiser, C.A. Identification of Sec36p, Sec37p, and Sec38p: components of yeast complex that contains Sec34p and Sec35p. Mol. Biol. Cell 13, 1484–1500 (2002).

    Article  CAS  Google Scholar 

  13. Suvorova, E.S., Duden, R. & Lupashin, V.V. The Sec34/Sec35p complex, a Ypt1p effector required for retrograde intra-Golgi trafficking, interacts with Golgi SNAREs and COPI vesicle coat proteins. J. Cell Biol. 157, 631–643 (2002).

    Article  CAS  Google Scholar 

  14. Luo, Z. & Gallwitz, D. Biochemical and genetic evidence for the involvement of yeast Ypt6-GTPase in protein retrieval to different Golgi compartments. J. Biol. Chem. 278, 791–799 (2003).

    Article  CAS  Google Scholar 

  15. Schuldiner, M. et al. Exploration of the function and organization of the yeast early secretory pathway through an epistatic miniarray profile. Cell 123, 507–519 (2005).

    Article  CAS  Google Scholar 

  16. Shestakova, A., Suvorova, E., Pavliv, O., Khaidakova, G. & Lupashin, V. Interaction of the conserved oligomeric Golgi complex with t-SNARE Syntaxin5a/Sed5 enhances intra-Golgi SNARE complex stability. J. Cell Biol. 179, 1179–1192 (2007).

    Article  CAS  Google Scholar 

  17. Zolov, S.N. & Lupashin, V.V. Cog3p depletion blocks vesicle-mediated Golgi retrograde trafficking in HeLa cells. J. Cell Biol. 168, 747–759 (2005).

    Article  CAS  Google Scholar 

  18. Giaever, G. et al. Functional profiling of the Saccharomyces cerevisiae genome. Nature 418, 387–391 (2002).

    Article  CAS  Google Scholar 

  19. Deutschbauer, A.M. et al. Mechanisms of haploinsufficiency revealed by genome-wide profiling in yeast. Genetics 169, 1915–1925 (2005).

    Article  CAS  Google Scholar 

  20. Fotso, P., Koryakina, Y., Pavliv, O., Tsiomenko, A.B. & Lupashin, V.V. Cog1p plays a central role in the organization of the yeast conserved oligomeric Golgi complex. J. Biol. Chem. 280, 27613–27623 (2005).

    Article  CAS  Google Scholar 

  21. Ungar, D., Oka, T., Vasile, E., Krieger, M. & Hughson, F.M. Subunit architecture of the conserved oligomeric Golgi complex. J. Biol. Chem. 280, 32729–32735 (2005).

    Article  CAS  Google Scholar 

  22. Ren, Y. et al. A structure-based mechanism for vesicle capture by the multisubunit tethering complex Dsl1. Cell 139, 1119–1129 (2009).

    Article  CAS  Google Scholar 

  23. Tripathi, A., Ren, Y., Jeffrey, P.D. & Hughson, F.M. Structural characterization of Tip20p and Dsl1p, subunits of the Dsl1p vesicle tethering complex. Nat. Struct. Mol. Biol. 16, 114–123 (2009).

    Article  CAS  Google Scholar 

  24. Cavanaugh, L.F. et al. Structural analysis of conserved oligomeric Golgi complex subunit 2. J. Biol. Chem. 282, 23418–23426 (2007).

    Article  CAS  Google Scholar 

  25. Richardson, B.C. et al. Structural basis for a human glycosylation disorder caused by mutation of the COG4 gene. Proc. Natl. Acad. Sci. USA 106, 13329–13334 (2009).

    Article  CAS  Google Scholar 

  26. Dong, G., Hutagalung, A.H., Fu, C., Novick, P. & Reinisch, K.M. The structures of exocyst subunit Exo70p and the Exo84p C-terminal domains reveal a common motif. Nat. Struct. Mol. Biol. 12, 1094–1100 (2005).

    Article  CAS  Google Scholar 

  27. Sivaram, M.V., Furgason, M.L., Brewer, D.N. & Munson, M. The structure of the exocyst subunit Sec6p defines a conserved architecture with diverse roles. Nat. Struct. Mol. Biol. 13, 555–556 (2006).

    Article  CAS  Google Scholar 

  28. Wu, S., Mehta, S.Q., Pichaud, F., Bellen, H.J. & Quiocho, F.A. Sec15 interacts with Rab11 via a novel domain and affects Rab11 localization in vivo. Nat. Struct. Mol. Biol. 12, 879–885 (2005).

    Article  CAS  Google Scholar 

  29. Scheich, C., Kummel, D., Soumailakakis, D., Heinemann, U. & Bussow, K. Vectors for co-expression of an unrestricted number of proteins. Nucleic Acids Res. 35, e43 (2007).

    Article  Google Scholar 

  30. Wu, X. et al. Mutation of the COG complex subunit gene COG7 causes a lethal congenital disorder. Nat. Med. 10, 518–523 (2004).

    Article  CAS  Google Scholar 

  31. Foulquier, F. et al. A new inborn error of glycosylation due to a Cog8 deficiency reveals a critical role for the Cog1-Cog8 interaction in COG complex formation. Hum. Mol. Genet. 16, 717–730 (2007).

    Article  CAS  Google Scholar 

  32. Kranz, C. et al. COG8 deficiency causes new congenital disorder of glycosylation type IIh. Hum. Mol. Genet. 16, 731–741 (2007).

    Article  CAS  Google Scholar 

  33. Foulquier, F. et al. Conserved oligomeric Golgi complex subunit 1 deficiency reveals a previously uncharacterized congenital disorder of glycosylation type II. Proc. Natl. Acad. Sci. USA 103, 3764–3769 (2006).

    Article  CAS  Google Scholar 

  34. Hsu, S.C. et al. Subunit composition, protein interactions, and structures of the mammalian brain sec6/8 complex and septin filaments. Neuron 20, 1111–1122 (1998).

    Article  CAS  Google Scholar 

  35. Sutton, R.B., Fasshauer, D., Jahn, R. & Brunger, A.T. Crystal structure of a SNARE complex involved in synaptic exocytosis at 2.4 Å resolution. Nature 395, 347–353 (1998).

    Article  CAS  Google Scholar 

  36. Reynders, E. et al. Golgi function and dysfunction in the first COG4-deficient CDG type II patient. Hum. Mol. Genet. 18, 3244–3256 (2009).

    Article  CAS  Google Scholar 

  37. Laufman, O., Kedan, A., Hong, W. & Lev, S. Direct interaction between the COG complex and the SM protein, Sly1, is required for Golgi SNARE pairing. EMBO J. 28, 2006–2017 (2009).

    Article  CAS  Google Scholar 

  38. Semenza, J.C., Hardwick, K.G., Dean, N. & Pelham, H.R. ERD2, a yeast gene required for the receptor-mediated retrieval of luminal ER proteins from the secretory pathway. Cell 61, 1349–1357 (1990).

    Article  CAS  Google Scholar 

  39. Voth, W.P., Jiang, Y.W. & Stillman, D.J. New 'marker swap' plasmids for converting selectable markers on budding yeast gene disruptions and plasmids. Yeast 20, 985–993 (2003).

    Article  CAS  Google Scholar 

  40. Longtine, M.S. et al. Additional modules for versatile and economical PCR-based gene deletion and modification in Saccharomyces cerevisiae. Yeast 14, 953–961 (1998).

    Article  CAS  Google Scholar 

  41. Rose, M.D., Misra, L.M. & Vogel, J.P. KAR2, a karyogamy gene, is the yeast homolog of the mammalian BiP/GRP78 gene. Cell 57, 1211–1221 (1989).

    Article  CAS  Google Scholar 

  42. Ohi, M., Li, Y., Cheng, Y. & Walz, T. Negative staining and image classification—powerful tools in modern electron microscopy. Biol. Proced. Online 6, 23–34 (2004).

    Article  CAS  Google Scholar 

  43. Li, Z., Hite, R.K., Cheng, Y. & Walz, T. Evaluation of imaging plates as recording medium for images of negatively stained single particles and electron diffraction patterns of two-dimensional crystals. J. Electron Microsc. (Tokyo) 59, 53–63 (2010).

    Article  CAS  Google Scholar 

  44. Ludtke, S.J., Baldwin, P.R. & Chiu, W. EMAN: semiautomated software for high-resolution single-particle reconstructions. J. Struct. Biol. 128, 82–97 (1999).

    Article  CAS  Google Scholar 

  45. Frank, J. et al. SPIDER and WEB: processing and visualization of images in 3D electron microscopy and related fields. J. Struct. Biol. 116, 190–199 (1996).

    Article  CAS  Google Scholar 

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Acknowledgements

We gratefully acknowledge M. Rose and C. Ydenberg (Princeton University) for providing strains, reagents, technical assistance and valuable advice, as well as S. Silverman and L. Schepis (Princeton University) for providing reagents and technical assistance. We also thank V. Lupashin, B. Richardson, D. Ungar and M. Paul for useful discussions. This work was supported by a grant to F.M.H. from the US National Institutes of Health (GM071574). C.K.Y. acknowledges fellowships from the Jane Coffin-Childs Memorial Fund and the Canadian Institutes for Health Research. T.W. is an investigator in the Howard Hughes Medical Institute.

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Author notes

  1. Joshua A Lees and Calvin K Yip: These authors contributed equally to this paper and are listed in alphabetical order.

Authors and Affiliations

  1. Department of Molecular Biology, Princeton University, Princeton, New Jersey, USA

    Joshua A Lees & Frederick M Hughson

  2. Department of Cell Biology, Harvard Medical School, Boston, Massachusetts, USA

    Calvin K Yip & Thomas Walz

  3. Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts, USA

    Thomas Walz

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  1. Joshua A Lees
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  4. Frederick M Hughson
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Contributions

J.A.L. conducted yeast experiments and prepared samples for EM. C.K.Y. conducted EM experiments and analyzed the data. All authors discussed the results and wrote the paper.

Corresponding author

Correspondence to Frederick M Hughson.

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Lees, J., Yip, C., Walz, T. et al. Molecular organization of the COG vesicle tethering complex. Nat Struct Mol Biol 17, 1292–1297 (2010). https://doi.org/10.1038/nsmb.1917

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