The membrane-associated proteins FCHo and SGIP are allosteric activators of the AP2 clathrin adaptor complex
- Stowers Institute for Medical Research, United States
- Howard Hughes Medical Institute, University of Utah, United States
- University of Utah, United States
- University of Washington, United States
- Ludwig-Maximilians-University, Germany
Abstract
The AP2 clathrin adaptor complex links protein cargo to the endocytic machinery but it is unclear how AP2 is activated on the plasma membrane. Here we demonstrate that the membrane-associated proteins FCHo and SGIP1 convert AP2 into an open, active conformation. We screened for C. elegans mutants that phenocopy the loss of AP2 subunits and found that AP2 remains inactive in fcho-1 mutants. A subsequent screen for bypass suppressors of fcho-1 nulls identified 71 compensatory mutations in all four AP2 subunits. Using a protease-sensitivity assay we show that these mutations restore the open conformation in vivo. The domain of FCHo that induces this rearrangement is not the F-BAR domain or the mu-homology domain, but rather is an uncharacterized 90 amino acid motif, found in both FCHo and SGIP proteins, that directly binds AP2. Thus, these proteins stabilize nascent endocytic pits by exposing membrane and cargo binding sites on AP2.
Article and author information
Author details
Reviewing Editor
- Suzanne R Pfeffer, Stanford University, United States
Version history
- Received: June 10, 2014
- Accepted: October 1, 2014
- Accepted Manuscript published: October 10, 2014 (version 1)
- Version of Record published: November 3, 2014 (version 2)
Copyright
© 2014, Hollopeter et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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The membrane-associated proteins FCHo and SGIP are allosteric activators of the AP2 clathrin adaptor complex
eLife 3:e03648.
https://doi.org/10.7554/eLife.03648
Further reading
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- Biochemistry and Chemical Biology
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Clathrin-mediated endocytosis is an evolutionarily ancient membrane transport system regulating cellular receptivity and responsiveness. Plasmalemma clathrin-coated structures range from unitary domed assemblies to expansive planar constructions with internal or flanking invaginated buds. Precisely how these morphologically-distinct coats are formed, and whether all are functionally equivalent for selective cargo internalization is still disputed. We have disrupted the genes encoding a set of early arriving clathrin-coat constituents, FCHO1 and FCHO2, in HeLa cells. Endocytic coats do not disappear in this genetic background; rather clustered planar lattices predominate and endocytosis slows, but does not cease. The central linker of FCHO proteins acts as an allosteric regulator of the prime endocytic adaptor, AP-2. By loading AP-2 onto the plasma membrane, FCHO proteins provide a parallel pathway for AP-2 activation and clathrin-coat fabrication. Further, the steady-state morphology of clathrin-coated structures appears to be a manifestation of the availability of the muniscin linker during lattice polymerization.
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