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Distinct and separable activities of the endocytic clathrin-coat components Fcho1/2 and AP-2 in developmental patterning

Nature Cell Biology volume 14, pages 488–501 (2012)Cite this article

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Abstract

Clathrin-mediated endocytosis occurs at multiple independent import sites on the plasma membrane, but how these positions are selected and how different cargo is simultaneously recognized is obscure. FCHO1 and FCHO2 are early-arriving proteins at surface clathrin assemblies and are speculated to act as compulsory coat nucleators, preceding the core clathrin adaptor AP-2. Here, we show that the μ-homology domain of FCHO1/2 represents an endocytic interaction hub. Translational silencing of fcho1 in zebrafish embryos causes strong dorsoventral patterning defects analogous to Bmp signal failure. The Fcho1 μ-homology domain interacts with the Bmp receptor Alk8, uncovering an endocytic component that positively modulates Bmp signal transmission. Still, the fcho1 morphant phenotype is distinct from severe embryonic defects apparent when AP-2 is depleted. Our data thus challenge the primacy of FCHO1/2 in coat initiation.

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Figure 1: Binding properties of FCHO1.
Figure 2: The μHD interaction hub.
Figure 3: Developmental defects of FCHO1 overexpression.
Figure 4: Fcho1 participates in early embryonic development.
Figure 5: The Fcho1–Alk8 association.
Figure 6: Fcho1 operates in a genetic Bmp-to-Smad signalling pathway.
Figure 7: AP-2 morphants are unlike fcho1/2 double morphants.
Figure 8: AP-2–clathrin coats persist in FCHO1 and FCHO2 siRNA-treated HeLa and BS-C-1 cells.

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Acknowledgements

We are indebted to our many colleagues for providing reagents that were essential for this study. Supported by National Institutes of Health grants R01 HL088016 (M.T.), R01 GM60979 (B.W.) and R01 DK53249 (L.M.T.).

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  1. Department of Cell Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15261, USA

    P. K. Umasankar, Subramaniam Sanker, James R. Thieman, Souvik Chakraborty & Linton M. Traub

  2. Department of Biology, The Johns Hopkins University, Baltimore, Maryland 21218, USA

    Beverly Wendland

  3. Department of Developmental Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15213, USA

    Michael Tsang

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Contributions

P.K.U., S.S., J.R.T., S.C. and L.M.T. designed, carried out and interpreted various experiments. B.W. and M.T. provided intellectual input, contributed to experimental design and advised on data interpretation. L.M.T. conceived and directed the overall project and wrote the manuscript with comments from all the authors.

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Correspondence to Linton M. Traub.

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Umasankar, P., Sanker, S., Thieman, J. et al. Distinct and separable activities of the endocytic clathrin-coat components Fcho1/2 and AP-2 in developmental patterning. Nat Cell Biol 14, 488–501 (2012). https://doi.org/10.1038/ncb2473

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