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ARFGAP1 promotes AP-2-dependent endocytosis

Nature Cell Biology volume 13, pages 559–567 (2011)Cite this article

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Abstract

COPI (coat protein I) and the clathrin–AP-2 (adaptor protein 2) complex are well-characterized coat proteins, but a component that is common to these two coats has not been identified. The GTPase-activating protein (GAP) for ADP-ribosylation factor 1 (ARF1), ARFGAP1, is a known component of the COPI complex. Here, we show that distinct regions of ARFGAP1 interact with AP-2 and coatomer (components of the COPI complex). Selectively disrupting the interaction of ARFGAP1 with either of these two coat proteins leads to selective inhibition in the corresponding transport pathway. The role of ARFGAP1 in AP-2-regulated endocytosis has mechanistic parallels with its roles in COPI transport, as both its GAP activity and coat function contribute to promoting AP-2 transport.

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Figure 1: Interactions with ARFGAP1 and effects of its knockdown.
Figure 2: Distinct requirements for ARFGAP1 binding to coatomer versus AP-2 and clathrin.
Figure 3: Disrupting the interaction between ARFGAP1 and either coatomer or AP-2 leads to selective disruption in transport pathways.
Figure 4: Surveying transport pathways affected by the depletion of ARFGAP1.
Figure 5: The role of ARFGAP1 in coated-pit formation.
Figure 6: ARFGAP1 and AP-2 interact specifically with surface TfR.
Figure 7: Characterizing the binding of TfR by ARFGAP1 and AP-2.
Figure 8: The GAP activity of ARFGAP1 promotes TfR endocytosis.

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Acknowledgements

We thank B. Wendland for critical comments. This work was financially supported by grants from the National Institutes of Health to V.W.H. (GM058615 and GM073016), T.K. (GM075252 and U54 AI057159—NERCE Imaging Resource) and J.F.C. (DK060065). A.L. was financially supported by the Telethon (Italy) and AIRC (Italy). H.G. was supported by a Marie Curie Fellowship. E.C. was supported by a GlaxoSmithKline fellowship.

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  1. Ming Bai, Helge Gad and Gabriele Turacchio: These authors contributed equally to this work

Authors and Affiliations

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

    Ming Bai, Jia-Shu Yang, Jian Li & Victor W. Hsu

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

    Helge Gad, Gabriele Turacchio, Galina V. Beznoussenko & Alberto Luini

  3. Department of Genetics, Microbiology and Toxicology, Stockholm University, 10691 Stockholm, Sweden

    Helge Gad

  4. Institute of Protein Biochemistry, National Research Council, Via Pietro, Castellino 111, 80131 Naples, Italy

    Gabriele Turacchio

  5. Department of Cell Biology, Harvard Medical School, and Immune Disease Institute, Boston, Massachusetts 02115, USA

    Emanuele Cocucci & Tomas Kirchhausen

  6. Department of Urology, University of Florida College of Medicine, Gainesville, Florida 32610, USA

    Zhongzhen Nie

  7. Laboratory of Cellular and Molecular Biology, National Cancer Institute, Bethesda, Maryland 20892, USA

    Ruibai Luo

  8. Department of Cell Biology, University of Alabama at Birmingham, Birmingham, Alabama 35294, USA

    Lianwu Fu & James F. Collawn

  9. Telethon Institute of Genetics and Medicine, Via Pietro Castellino 111, 80131 Napoli, Italy

    Alberto Luini

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Contributions

M.B., H.G., G.T., E.C., J-S.Y., J.L., G.V.B., Z.N., L.F. and R.L. carried out experiments and data analyses. V.W.H., A.L., T.K. and J.F.C. supervised the work. V.W.H., A.L., H.G. and M.B. wrote the manuscript.

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Correspondence to Victor W. Hsu.

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Bai, M., Gad, H., Turacchio, G. et al. ARFGAP1 promotes AP-2-dependent endocytosis. Nat Cell Biol 13, 559–567 (2011). https://doi.org/10.1038/ncb2221

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