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Structural characterization of Tip20p and Dsl1p, subunits of the Dsl1p vesicle tethering complex

Nature Structural & Molecular Biology volume 16, pages 114–123 (2009)Cite this article

Abstract

Multisubunit tethering complexes are essential for intracellular trafficking and have been proposed to mediate the initial interaction between vesicles and the membranes with which they fuse. Here we report initial structural characterization of the Dsl1p complex, whose three subunits are essential for trafficking from the Golgi apparatus to the endoplasmic reticulum (ER). Crystal structures reveal that two of the three subunits, Tip20p and Dsl1p, resemble known subunits of the exocyst complex, establishing a structural connection among several multisubunit tethering complexes and implying that many of their subunits are derived from a common progenitor. We show, moreover, that Tip20p and Dsl1p interact directly via N-terminal α-helices. Finally, we establish that different Dsl1p complex subunits bind independently to different ER SNARE proteins. Our results map out two alternative protein-interaction networks capable of tethering COPI-coated vesicles, via the Dsl1p complex, to ER membranes.

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Figure 1: X-ray crystal structures of S. cerevisiae Dsl1p complex subunits.
Figure 2: The Tip20p and Dsl1p subunits of the Dsl1p complex form stoichiometric heterodimers.
Figure 3: Structural and biochemical characterization of the Tip20p–Dsl1p interaction.
Figure 4: Reconstitution of the heterotrimeric Dsl1p complex.
Figure 5: ER SNAREs Sec20p and Use1p bind Dsl1p complex via different subunits.
Figure 6: Schematic model for the tethering of Golgi-derived retrograde trafficking vesicles to the ER via bivalent attachment of the Dsl1p complex to the ER SNAREs Use1p and Sec20p.

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Acknowledgements

We gratefully acknowledge B. Kokona and R. Fairman for sedimentation velocity analytical ultracentrifugation; the staff of the National Synchrotron Light Source X25 and X29 beamlines for assistance with X-ray data collection; O. Perisic for advice on crystallization; M. Diefenbacher and A. Spang for many fruitful discussions and for communicating results before publication; K. Büssow (Max Planck Institute for Molecular Genetics, Berlin) for reagents; and S. Munro, M. Munson, A. Spang and members of the Hughson laboratory for critical comments on the manuscript. This work was supported by the US National Institutes of Health grant GM071574.

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  1. Arati Tripathi and Yi Ren: These authors contributed equally to this work.

Authors and Affiliations

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

    Arati Tripathi, Yi Ren, Philip D Jeffrey & Frederick M Hughson

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Correspondence to Frederick M Hughson.

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Tripathi, A., Ren, Y., Jeffrey, P. et al. Structural characterization of Tip20p and Dsl1p, subunits of the Dsl1p vesicle tethering complex. Nat Struct Mol Biol 16, 114–123 (2009). https://doi.org/10.1038/nsmb.1548

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