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Structure of the yeast polarity protein Sro7 reveals a SNARE regulatory mechanism

Nature volume 446, pages 567–571 (2007)Cite this article

Abstract

Polarized exocytosis requires coordination between the actin cytoskeleton and the exocytic machinery responsible for fusion of secretory vesicles at specific sites on the plasma membrane1. Fusion requires formation of a complex between a vesicle-bound R-SNARE and plasma membrane Qa, Qb and Qc SNARE proteins2. Proteins in the lethal giant larvae protein family, including lethal giant larvae and tomosyn in metazoans and Sro7 in yeast, interact with Q-SNAREs and are emerging as key regulators of polarized exocytosis3. The crystal structure of Sro7 reveals two seven-bladed WD40 β-propellers followed by a 60-residue-long ‘tail’, which is bound to the surface of the amino-terminal propeller. Deletion of the Sro7 tail enables binding to the Qbc SNARE region of Sec9 and this interaction inhibits SNARE complex assembly. The N-terminal domain of Sec9 provides a second, high-affinity Sro7 interaction that is unaffected by the tail. The results suggest that Sro7 acts as an allosteric regulator of exocytosis through interactions with factors that control the tail. Sequence alignments indicate that lethal giant larvae and tomosyn have a two-β-propeller fold similar to that of Sro7, but only tomosyn appears to retain the regulatory tail.

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Figure 1: Overview of the Sro7 structure.
Figure 2: Surface representation of Sro7, with residues coloured from white to yellow by increasing conservation.
Figure 3: Interaction of Sec9 with Sro7.
Figure 4: Proposed model for how Sro7 coordinates release of Sec9 with arrival of a secretory vesicle.

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References

  1. Pruyne, D., Legesse-Miller, A., Gao, L., Dong, Y. & Bretscher, A. Mechanisms of polarized growth and organelle segregation in yeast. Annu. Rev. Cell Dev. Biol. 20, 559–591 (2004)

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  3. Wirtz-Peitz, F. & Knoblich, J. A. Lethal giant larvae take on a life of their own. Trends Cell Biol. 16, 234–241 (2006)

    Article  CAS  Google Scholar 

  4. Lehman, K., Rossi, G., Adamo, J. E. & Brennwald, P. Yeast homologues of tomosyn and lethal giant larvae function in exocytosis and are associated with the plasma membrane SNARE, Sec9. J. Cell Biol. 146, 125–140 (1999)

    CAS  PubMed  PubMed Central  Google Scholar 

  5. Zhang, X. et al. Lethal giant larvae proteins interact with the exocyst complex and are involved in polarized exocytosis. J. Cell Biol. 170, 273–283 (2005)

    Article  CAS  Google Scholar 

  6. Grosshans, B. L. et al. The yeast lgl family member Sro7p is an effector of the secretory Rab GTPase Sec4p. J. Cell Biol. 172, 55–66 (2006)

    Article  CAS  Google Scholar 

  7. Adamo, J. E., Rossi, G. & Brennwald, P. The Rho GTPase Rho3 has a direct role in exocytosis that is distinct from its role in actin polarity. Mol. Biol. Cell 10, 4121–4133 (1999)

    Article  CAS  Google Scholar 

  8. Adamo, J. E. et al. Yeast Cdc42 functions at a late step in exocytosis, specifically during polarized growth of the emerging bud. J. Cell Biol. 155, 581–592 (2001)

    Article  CAS  Google Scholar 

  9. Kagami, M., Toh-e, A. & Matsui, Y. Sro7p, a Saccharomyces cerevisiae counterpart of the tumor suppressor l(2)gl protein, is related to myosins in function. Genetics 149, 1717–1727 (1998)

    CAS  PubMed  PubMed Central  Google Scholar 

  10. Vasioukhin, V. Lethal giant puzzle of Lgl. Dev. Neurosci. 28, 13–24 (2006)

    Article  CAS  Google Scholar 

  11. Gracheva, E. O. et al. Tomosyn inhibits synaptic vesicle priming in Caenorhabditis elegans. PLoS Biol. 4, e261 (2006)

    Article  Google Scholar 

  12. McEwen, J. M., Madison, J. M., Dybbs, M. & Kaplan, J. M. Antagonistic regulation of synaptic vesicle priming by Tomosyn and UNC-13. Neuron 51, 303–315 (2006)

    Article  CAS  Google Scholar 

  13. Musch, A. et al. Mammalian homolog of Drosophila tumor suppressor lethal (2) giant larvae interacts with basolateral exocytic machinery in Madin-Darby canine kidney cells. Mol. Biol. Cell 13, 158–168 (2002)

    Article  CAS  Google Scholar 

  14. Hatsuzawa, K., Lang, T., Fasshauer, D., Bruns, D. & Jahn, R. The R-SNARE motif of tomosyn forms SNARE core complexes with syntaxin 1 and SNAP-25 and down-regulates exocytosis. J. Biol. Chem. 278, 31159–31166 (2003)

    Article  CAS  Google Scholar 

  15. Pobbati, A. V., Razeto, A., Boddener, M., Becker, S. & Fasshauer, D. Structural basis for the inhibitory role of tomosyn in exocytosis. J. Biol. Chem. 279, 47192–47200 (2004)

    Article  CAS  Google Scholar 

  16. Mohri, K., Vorobiev, S., Fedorov, A. A., Almo, S. C. & Ono, S. Identification of functional residues on Caenorhabditis elegans actin-interacting protein 1 (UNC-78) for disassembly of actin depolymerizing factor/cofilin-bound actin filaments. J. Biol. Chem. 279, 31697–31707 (2004)

    Article  CAS  Google Scholar 

  17. Voegtli, W. C., Madrona, A. Y. & Wilson, D. K. The structure of Aip1p, a WD repeat protein that regulates cofilin-mediated actin depolymerization. J. Biol. Chem. 278, 34373–34379 (2003)

    Article  CAS  Google Scholar 

  18. Betschinger, J., Eisenhaber, F. & Knoblich, J. A. Phosphorylation-induced autoinhibition regulates the cytoskeletal protein Lethal (2) giant larvae. Curr. Biol. 15, 276–282 (2005)

    Article  CAS  Google Scholar 

  19. Baba, T., Sakisaka, T., Mochida, S. & Takai, Y. PKA-catalyzed phosphorylation of tomosyn and its implication in Ca2+-dependent exocytosis of neurotransmitter. J. Cell Biol. 170, 1113–1125 (2005)

    Article  CAS  Google Scholar 

  20. Brennwald, P. et al. Sec9 is a SNAP-25-like component of a yeast SNARE complex that may be the effector of Sec4 function in exocytosis. Cell 79, 245–258 (1994)

    Article  CAS  Google Scholar 

  21. Fasshauer, D., Sutton, R. B., Brunger, A. T. & Jahn, R. Conserved structural features of the synaptic fusion complex: SNARE proteins reclassified as Q- and R-SNAREs. Proc. Natl Acad. Sci. USA 95, 15781–15786 (1998)

    Article  ADS  CAS  Google Scholar 

  22. Goud, B., Salminen, A., Walworth, N. C. & Novick, P. J. A GTP-binding protein required for secretion rapidly associates with secretory vesicles and the plasma membrane in yeast. Cell 53, 753–768 (1988)

    Article  CAS  Google Scholar 

  23. Boyd, C., Hughes, T., Pypaert, M. & Novick, P. Vesicles carry most exocyst subunits to exocytic sites marked by the remaining two subunits, Sec3p and Exo70p. J. Cell Biol. 167, 889–901 (2004)

    Article  CAS  Google Scholar 

  24. Gari, E., Piedrafita, L., Aldea, M. & Herrero, E. A set of vectors with a tetracycline-regulatable promoter system for modulated gene expression in Saccharomyces cerevisiae. Yeast 13, 837–848 (1997)

    Article  CAS  Google Scholar 

  25. Baker, N. A., Sept, D., Joseph, S., Holst, M. J. & McCammon, J. A. Electrostatics of nanosystems: application to microtubules and the ribosome. Proc. Natl Acad. Sci. USA 98, 10037–10041 (2001)

    Article  ADS  CAS  Google Scholar 

  26. Landau, M. et al. ConSurf 2005: the projection of evolutionary conservation scores of residues on protein structures. Nucleic Acids Res. 33, W299–W302 (2005)

    Article  ADS  CAS  Google Scholar 

  27. Nicholson, K. L. et al. Regulation of SNARE complex assembly by an N-terminal domain of the t-SNARE Sso1p. Nature Struct. Biol. 5, 793–802 (1998)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank L. Katz for help with the yeast two-hybrid analysis, and S. Kaiser for assistance with mass spectrometry. Diffraction data were measured at the Advanced Light Source and the Stanford Synchrotron Radiation Laboratory. D.A.H. was supported by a fellowship from the American Cancer Society. This work was supported by NIH grants to P.J.B. and W.I.W and a grant from the G. Harold and Leila Y. Mathers Foundation to P.J.B.

Author Contributions Crystallographic and biochemical experiments were designed by D.A.H. and W.I.W. and performed by D.A.H. Plasmid and strain construction were designed and performed by D.A.H. and P.J.B. Yeast two-hybrid analysis, secretion assays and analysis of mutant phenotypes were designed by P.J.B. and performed by A.A. and A.G. D.A.H. and W.I.W. wrote the paper and all authors made editorial comments.

Coordinates and structure factors have been deposited in the Protein Data Bank under accession number 2OAJ.

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Authors and Affiliations

  1. Departments of Structural Biology and of Molecular and Cellular Physiology, Stanford University School of Medicine, 299 Campus Drive West, Stanford, California 94305-5126, USA,

    Douglas A. Hattendorf & William I. Weis

  2. Department of Cell and Developmental Biology, University of North Carolina, 538 Taylor Hall, Chapel Hill, North Carolina, 27599-7090, USA,

    Anna Andreeva, Akanksha Gangar & Patrick J. Brennwald

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Correspondence to Patrick J. Brennwald or William I. Weis.

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Hattendorf, D., Andreeva, A., Gangar, A. et al. Structure of the yeast polarity protein Sro7 reveals a SNARE regulatory mechanism. Nature 446, 567–571 (2007). https://doi.org/10.1038/nature05635

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