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Phox domain interaction with PtdIns(3)P targets the Vam7 t-SNARE to vacuole membranes

Nature Cell Biology volume 3, pages 613–618 (2001)Cite this article

An Erratum to this article was published on 01 July 2001

Abstract

Specific recognition of phosphoinositides is crucial for protein sorting and membrane trafficking. Protein transport to the yeast vacuole depends on the Vam7 t-SNARE and its phox homology (PX) domain. Here, we show that the PX domain of Vam7 targets to vacuoles in vivo in a manner dependent on phosphatidylinositol 3-phosphate generation. A novel phosphatidylinositol-3-phosphate-binding motif and an exposed loop that interacts with the lipid bilayer are identified by nuclear magnetic resonance spectroscopy. Conservation of key structural and binding site residues across the diverse PX family indicates a shared fold and phosphoinositide recognition function.

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Figure 1: Phosphatidylinositol 3-phosphate-dependent localization of GFP–Vam7 by fluorescence microscopy.
Figure 2: Specific phosphatidylinositol 3-phosphate (PtdIns(3)P) recognition by the Vam7 phox homology (PX) domain.
Figure 3: Identification of residues involved in specific phosphatidylinositol 3-phosphate (PtdIns(3)P) ligation.
Figure 4: Lipid micelle interactions of the Vam7 phox homology (PX) domain.
Figure 5: Alignment of phox homology (PX) domain sequences.

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Acknowledgements

We thank D. G. S. Capelluto, J. L. Enmon, D. N. M. Jones and A. E. Wurmser for discussions and L. E. Kay for pulse sequences. We also thank C. Sette, P. Iaquinta, J. Thorner, X. Song, W. Xu, A. Zhang, G. Huang, X. Liang, J. V. Virbasius, M. P. Czech and G. W. Zhou for sharing unpublished data. We are supported by the Pew Scholars Program, University of Colorado Cancer Center and University of Colorado Health Sciences Center NMR Facility.

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  1. Matthew L. Cheever and Trey K. Sato: These authors contributed equally to this work

Authors and Affiliations

  1. Molecular Biology Program, University of Colorado Health Sciences Center, 4200 East Ninth Avenue, Denver, 80262, Colorado, USA

    Matthew L. Cheever

  2. Department of Pharmacology, University of Colorado Health Sciences Center, 4200 East Ninth Avenue, Denver, 80262, Colorado, USA

    Tonny de Beer, Tatiana G. Kutateladze & Michael Overduin

  3. Division of Cellular and Molecular Medicine and Howard Hughes Medical Institute, University of California at San Diego, School of Medicine, La Jolla, 92093, California, USA

    Trey K. Sato & Scott D. Emr

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Correspondence to Michael Overduin.

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Cheever, M., Sato, T., de Beer, T. et al. Phox domain interaction with PtdIns(3)P targets the Vam7 t-SNARE to vacuole membranes. Nat Cell Biol 3, 613–618 (2001). https://doi.org/10.1038/35083000

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