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Molecular architecture of the TRAPPII complex and implications for vesicle tethering

Nature Structural & Molecular Biology volume 17, pages 1298–1304 (2010)Cite this article

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Abstract

Multisubunit tethering complexes participate in the process of vesicle tethering—the initial interaction between transport vesicles and their acceptor compartments. TRAPPII (named for transport protein particle II) is a highly conserved tethering complex that functions in the late Golgi apparatus and consists of all of the subunits of TRAPPI and three additional, specific subunits. We have purified native yeast TRAPPII and characterized its structure and subunit organization by single-particle EM. Our data show that the nine TRAPPII components form a core complex that dimerizes into a three-layered, diamond-shaped structure. The TRAPPI subunits assemble into TRAPPI complexes that form the outer layers. The three TRAPPII-specific subunits cap the ends of TRAPPI and form the middle layer, which is responsible for dimerization. TRAPPII binds the Ypt1 GTPase and probably uses the TRAPPI catalytic core to promote guanine nucleotide exchange. We discuss the implications of the structure of TRAPPII for coat interaction and TRAPPII-associated human pathologies.

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Figure 1: Purification and negative stain EM analysis of yeast TRAPPII.
Figure 2: Three-dimensional reconstructions of TRAPPII.
Figure 3: The TRAPPI subunits localize to the periphery of TRAPPII.
Figure 4: Localization of Trs120 and Trs130.
Figure 5: Kre11 mediates dimerization of TRAPPII.
Figure 6: TRAPPII binds Ypt1.

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Acknowledgements

We thank J. Nicols for technical assistance with purification and data processing and Z. Li for assistance with microscopy and image processing. C.K.Y. acknowledges fellowships from the Jane Coffin Childs Memorial Fund and the Canadian Institutes of Health Research. T.W. is an investigator in the Howard Hughes Medical Institute. The molecular EM facility at Harvard Medical School was established with a generous donation from the Giovanni Armenise Harvard Center for Structural Biology and is maintained with funds from US National Institutes of Health (NIH) grant PO1 GM62580 (to S.C. Harrison). Molecular graphics images were produced using the UCSF Chimera package from the Resource for Biocomputing, Visualization, and Informatics at the University of California, San Francisco (supported by NIH P41 RR-01081).

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Authors and Affiliations

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

    Calvin K Yip, Julia Berscheminski & Thomas Walz

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

    Thomas Walz

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  1. Calvin K Yip
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Contributions

C.K.Y. and T.W. conceived the experiments. C.K.Y. performed all experiments. J.B. assisted in generating GFP-labeled yeast strains and data processing. C.K.Y. and T.W. analyzed the data and wrote the manuscript.

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Correspondence to Calvin K Yip.

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Yip, C., Berscheminski, J. & Walz, T. Molecular architecture of the TRAPPII complex and implications for vesicle tethering. Nat Struct Mol Biol 17, 1298–1304 (2010). https://doi.org/10.1038/nsmb.1914

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