Format

Send to

Choose Destination
Dev Cell. 2019 Aug 19;50(4):494-508.e11. doi: 10.1016/j.devcel.2019.07.017.

Temporal Ordering in Endocytic Clathrin-Coated Vesicle Formation via AP2 Phosphorylation.

Author information

1
CIMR, WT/MRC Building, Hills Road, Cambridge CB2 0QQ, UK.
2
CIMR, WT/MRC Building, Hills Road, Cambridge CB2 0QQ, UK. Electronic address: zk241@cam.ac.uk.
3
Czech Academy of Sciences, Institute of Information Theory and Automation, Pod Vodarenskou vezi 4, 182 08 Prague 8, Czech Republic.
4
MRC Laboratory of Molecular Biology, Cambridge Biomedical Campus, Francis Crick Avenue, Cambridge CB2 0QH, UK.
5
University of Grenoble Alpes, CNRS, CEA, IBS, 38000 Grenoble, France.
6
The Francis Crick Institute, 1 Midland Road, London NW1 1ST, UK.
7
Center for Molecular Medicine (CMMC), University of Cologne, Robert-Koch-Straße 21, 50931 Cologne, Germany.
8
Institute for Biochemistry I, Medical Faulty, University of Cologne, Joseph-Stelzmann-Straße 52, 50931 Cologne, Germany. Electronic address: shoening@uni-koeln.de.

Abstract

Clathrin-mediated endocytosis (CME) is key to maintaining the transmembrane protein composition of cells' limiting membranes. During mammalian CME, a reversible phosphorylation event occurs on Thr156 of the μ2 subunit of the main endocytic clathrin adaptor, AP2. We show that this phosphorylation event starts during clathrin-coated pit (CCP) initiation and increases throughout CCP lifetime. μ2Thr156 phosphorylation favors a new, cargo-bound conformation of AP2 and simultaneously creates a binding platform for the endocytic NECAP proteins but without significantly altering AP2's cargo affinity in vitro. We describe the structural bases of both. NECAP arrival at CCPs parallels that of clathrin and increases with μ2Thr156 phosphorylation. In turn, NECAP recruits drivers of late stages of CCP formation, including SNX9, via a site distinct from where NECAP binds AP2. Disruption of the different modules of this phosphorylation-based temporal regulatory system results in CCP maturation being delayed and/or stalled, hence impairing global rates of CME.

KEYWORDS:

AAK1; AP2 endocytic adaptor; NECAP; NMR; Numb-associated kinases (NAK); SNX9; TIRF; clathrin-mediated endocytosis; crystallography; regulation by phosphorylation

Supplemental Content

Full text links

Icon for Elsevier Science Icon for PubMed Central
Loading ...
Support Center