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Interactions within the yeast t-SNARE Sso1p that control SNARE complex assembly

Nature Structural Biology volume 7, pages 894–902 (2000)Cite this article

Abstract

In the eukaryotic secretory and endocytic pathways, transport vesicles shuttle cargo among intracellular organelles and to and from the plasma membrane. Cargo delivery entails fusion of the transport vesicle with its target, a process thought to be mediated by membrane bridging SNARE protein complexes. Temporal and spatial control of intracellular trafficking depends in part on regulating the assembly of these complexes. In vitro, SNARE assembly is inhibited by the closed conformation adopted by the syntaxin family of SNAREs. To visualize this closed conformation directly, the X-ray crystal structure of a yeast syntaxin, Sso1p, has been determined and refined to 2.1 Å resolution. Mutants designed to destabilize the closed conformation exhibit accelerated rates of SNARE assembly. Our results provide insight into the mechanism of SNARE assembly and its intramolecular and intermolecular regulation.

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Figure 1: The N-terminal domain of Sso1p is required for cell viability.
Figure 2: The stability of Sso1p and the rate of binary SNARE complex assembly are inversely dependent on pH.
Figure 3: Sso1p structure.
Figure 4: Comparison between Sso1p and syntaxin–nSec1 structures.
Figure 5: Stereo views of buried waters and hydrophobic clusters in Sso1p.
Figure 6: Superposition of Sso1p and the neuronal SNARE core complex.
Figure 7: Mutagenic analysis of SNARE assembly.

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Acknowledgements

We thank J. Carey, C. Carr, L. Cavanaugh, E. Grote, R. Miller, S. Miller, P. Novick, B. Reilly, M. Rose, Y. Shi, and G. Waters for gifts of materials and for helpful advice. We are particularly grateful to J. Lerman for guidance in designing mutants and assistance in X-ray data collection. This work was supported by the American Heart Association (M.M.), the Searle Scholars and Beckman Young Investigators programs (F.M.H.), and the N.I.H. (M.M. and F.M.H.).

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  1. Mary Munson, Xin Chen, Amy E. Cocina, Susan M. Schultz and Frederick M. Hughson: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Molecular Biology, Princeton University, Princeton, 08544, New Jersey, USA

    Mary Munson, Xin Chen, Amy E. Cocina, Susan M. Schultz & Frederick M. Hughson

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Correspondence to Frederick M. Hughson.

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Munson, M., Chen, X., Cocina, A. et al. Interactions within the yeast t-SNARE Sso1p that control SNARE complex assembly. Nat Struct Mol Biol 7, 894–902 (2000). https://doi.org/10.1038/79659

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