PT  - JOURNAL ARTICLE
AU  - Gwendolyn M Beacham
AU  - Edward A Partlow
AU  - Jeffrey J Lange
AU  - Gunther Hollopeter
TI  - NECAPs are negative regulators of the AP2 clathrin adaptor complex
AID  - 10.1101/192898
DP  - 2017 Jan 01
TA  - bioRxiv
PG  - 192898
4099  - http://biorxiv.org/content/early/2017/09/22/192898.short
4100  - http://biorxiv.org/content/early/2017/09/22/192898.full
AB  - Eukaryotic cells internalize transmembrane receptors via clathrin-mediated endocytosis, but it remains unclear how the machinery underpinning this process is regulated. We recently discovered that membrane-associated muniscin proteins such as FCHo and SGIP initiate endocytosis by converting the AP2 clathrin adaptor complex to an open, active conformation that is then phosphorylated (Hollopeter et al., 2014). Here we report that loss of ncap-1, the sole C. elegans gene encoding an adaptiN Ear-binding Coat-Associated Protein (NECAP), bypasses the requirement for FCHO-1. Biochemical analyses reveal AP2 accumulates in an open, phosphorylated state in ncap-1 mutant worms, suggesting NECAPs promote the closed, inactive conformation of AP2. Consistent with this model, NECAPs preferentially bind open and phosphorylated forms of AP2 in vitro and localize with constitutively open AP2 mutants in vivo. NECAPs do not associate with phosphorylation-defective AP2 mutants, implying that phosphorylation precedes NECAP recruitment. We propose NECAPs function late in endocytosis to inactivate AP2.