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A role for phosphatidic acid in COPI vesicle fission yields insights into Golgi maintenance

Nature Cell Biology volume 10, pages 1146–1153 (2008)Cite this article

Abstract

Proteins essential for vesicle formation by the Coat Protein I (COPI) complex are being identified, but less is known about the role of specific lipids. Brefeldin-A ADP-ribosylated substrate (BARS) functions in the fission step of COPI vesicle formation. Here, we show that BARS induces membrane curvature in cooperation with phosphatidic acid. This finding has allowed us to further delineate COPI vesicle fission into two sub-stages: 1) an earlier stage of bud-neck constriction, in which BARS and other COPI components are required, and 2) a later stage of bud-neck scission, in which phosphatidic acid generated by phospholipase D2 (PLD2) is also required. Moreover, in contrast to the disruption of the Golgi seen on perturbing the core COPI components (such as coatomer), inhibition of PLD2 causes milder disruptions, suggesting that such COPI components have additional roles in maintaining Golgi structure other than through COPI vesicle formation.

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Figure 1: Liposome tubulation by BARS requires phosphatidic acid (PA).
Figure 2: Perturbation of phosphatidic acid generated by PLD2 affects COPI transport.
Figure 3: COPI vesicle formation requires phosphatidic acid generated by PLD2.
Figure 4: PLD2 is required for the late stage of COPI vesicle fission.
Figure 5: Golgi morphology and COPI distribution upon PLD2 depletion.

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Acknowledgements

We thank Jian Li and Ming Bai for advice and discussions, and Richard Premont for technical assistance. This work was funded by grants from the NIH to V.W.H. (GM058615), M.A.F. (GM071520) and G.D. (GM071475), from Telethon (Italy), AIRC (Italy) to A.L. and A.M., and from the Canadian Institutes of Health Research (S.B.). H.G. was supported by a Marie Curie Fellowship.

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Authors and Affiliations

  1. Division of Rheumatology, and Department of Medicine, Immunology and Allergy, Brigham and Women's Hospital, Harvard Medical School, Boston, 02115, MA, USA

    Jia-Shu Yang, Stella Y. Lee, Leiliang Zhang & Victor W. Hsu

  2. Department of Cell Biology and Oncology, Consorzio Mario Negri Sud, 66030, Santa Maria Imbaro (Chieti), Italy

    Helge Gad, Alexander Mironov, Galina V. Beznoussenko, Carmen Valente, Gabriele Turacchio & Alberto Luini

  3. Department of Pharmacology and the Center for Developmental Genetics, Stony Brook University, Stony Brook, 11794, NY, USA

    Akua N. Bonsra, Guangwei Du & Michael A. Frohman

  4. Department of Clinical and Experimental Medicine, Università del Piemonte Orientale, 28100, Novara, Italy

    Gianluca Baldanzi & Andrea Graziani

  5. Le Centre Hospitalier Universitaire de Quebec, Pavillon CHUL, Rhumatologie et Immunology, Quebec, G1V4G2, Canada

    Sylvain Bourgoin

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Contributions

Experiments were performed mainly by J.S.Y., with help from H.D., S.Y.L., A.M., L.Z., G.V.B., C.V., G.T., A.N.B., G.D., G.B., A.G. and S.B. The project was planned mainly by V.W.H. with help from A.L. and M.A.H. All authors participated in data analysis.

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Correspondence to Victor W. Hsu.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Information

Supplementary Figures S1, S2, S3, S4, S5, S6, S7, S8, S9 and Supplementary Table 1 (PDF 1100 kb)

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Yang, JS., Gad, H., Lee, S. et al. A role for phosphatidic acid in COPI vesicle fission yields insights into Golgi maintenance. Nat Cell Biol 10, 1146–1153 (2008). https://doi.org/10.1038/ncb1774

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