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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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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.

Author information

Affiliations

  1. Division of Rheumatology, Immunology and Allergy, Brigham and Women's Hospital, and Department of Medicine, Harvard Medical School, Boston, MA 02115 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, NY 11794, 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, Canada G1V4G2.

    • 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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The authors declare no competing financial interests.

Corresponding author

Correspondence to Victor W. Hsu.

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    Supplementary Information

    Supplementary Figures S1, S2, S3, S4, S5, S6, S7, S8, S9 and Supplementary Table 1

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DOI

https://doi.org/10.1038/ncb1774