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Topological restriction of SNARE-dependent membrane fusion

Nature volume 407, pages 194198 (14 September 2000) | Download Citation

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Abstract

To fuse transport vesicles with target membranes, proteins of the SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptors) complex must be located on both the vesicle (v-SNARE) and the target membrane (t-SNARE)1. In yeast, four integral membrane proteins, Sed5, Bos1, Sec22 and Bet1 (refs 2, 3,4,5,6), each probably contribute a single helix to form the SNARE complex that is needed for transport from endoplasmic reticulum to Golgi7,8,9,10,11. This generates a four-helix bundle12, which ultimately mediates the actual fusion event13. Here we explore how the anchoring arrangement of the four helices affects their ability to mediate fusion. We reconstituted two populations of phospholipid bilayer vesicles, with the individual SNARE proteins distributed in all possible combinations between them. Of the eight non-redundant permutations of four subunits distributed over two vesicle populations, only one results in membrane fusion. Fusion only occurs when the v-SNARE Bet1 is on one membrane and the syntaxin heavy chain Sed5 and its two light chains, Bos1 and Sec22, are on the other membrane where they form a functional t-SNARE. Thus, each SNARE protein is topologically restricted by design to function either as a v-SNARE or as part of a t-SNARE complex.

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Acknowledgements

We would like to thank R. J. Johnston for technical support. Research was supported by an NIH grant (to J.E.R.) and postdoctoral fellowships of the Medical Research Council of Canada (F.P.), the NIH (J.M.) and the Japanese Society for the Promotion of Science (R.F.).

Author information

Author notes

    • James A. McNew
    •  & Ryouichi Fukuda

    Present address: Department of Biochemistry and Cell Biology, Rice University, MS140, 6100 Main Street, PO Box 1892, Houston, Texas 77005, USA (J.A.M); Department of Biotechnology, University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan (R.F.).

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  1. Cellular Biochemistry and Biophysics Program, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, Box251 New York , New York 10021, USA

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Correspondence to James E. Rothman.

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https://doi.org/10.1038/35025076

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