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Dev Cell. 2016 Jun 6;37(5):428-43. doi: 10.1016/j.devcel.2016.05.003. Epub 2016 May 26.

Transient Fcho1/2⋅Eps15/R⋅AP-2 Nanoclusters Prime the AP-2 Clathrin Adaptor for Cargo Binding.

Author information

1
Department of Cell Biology, University of Pittsburgh School of Medicine, 3500 Terrace Street, S312 BST, Pittsburgh, PA 15261, USA.
2
Cambridge Institute for Medical Research, University of Cambridge, Cambridge CB2 0XY, UK.
3
Department of Cell Biology, University of Pittsburgh School of Medicine, 3500 Terrace Street, S312 BST, Pittsburgh, PA 15261, USA. Electronic address: traub@pitt.edu.

Abstract

Clathrin-coated vesicles form by rapid assembly of discrete coat constituents into a cargo-sorting lattice. How the sequential phases of coat construction are choreographed is unclear, but transient protein-protein interactions mediated by short interaction motifs are pivotal. We show that arrayed Asp-Pro-Phe (DPF) motifs within the early-arriving endocytic pioneers Eps15/R are differentially decoded by other endocytic pioneers Fcho1/2 and AP-2. The structure of an Eps15/R⋅Fcho1 μ-homology domain complex reveals a spacing-dependent DPF triad, bound in a mechanistically distinct way from the mode of single DPF binding to AP-2. Using cells lacking FCHO1/2 and with Eps15 sequestered from the plasma membrane, we establish that without these two endocytic pioneers, AP-2 assemblies are fleeting and endocytosis stalls. Thus, distinct DPF-based codes within the unstructured Eps15/R C terminus direct the assembly of temporary Fcho1/2⋅Eps15/R⋅AP-2 ternary complexes to facilitate conformational activation of AP-2 by the Fcho1/2 interdomain linker to promote AP-2 cargo engagement.

PMID:
27237791
PMCID:
PMC4921775
DOI:
10.1016/j.devcel.2016.05.003
[Indexed for MEDLINE]
Free PMC Article

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