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Temporal ordering in endocytic clathrin-coated vesicle formation via AP2 phosphorylation

DOI: 10.1016/j.devcel.2019.07.017 DOI Help

Authors: Antoni G. Wrobel (Cambridge Institute for Medical Research (CIMR)) , Zuzana Kadlecova (Cambridge Institute for Medical Research (CIMR)) , Jan Kamenicky (Czech Academy of Sciences) , Ji-chun Yang (MRC Laboratory of Molecular Biology, University of Cambridge) , Torsten Herrmann (University of Grenoble Alpes, CNRS, CEA, IBS) , Bernard T. Kelly (Cambridge Institute for Medical Research (CIMR)) , Airlie J. Mccoy (Cambridge Institute for Medical Research (CIMR)) , Philip R. Evans (MRC Laboratory of Molecular Biology, University of Cambridge) , Stephen Martin (The Francis Crick Institute) , Stefan Müller (Center for Molecular Medicine (CMMC), University of Cologne) , Filip Sroubek (Czech Academy of Sciences) , David Neuhaus (MRC Laboratory of Molecular Biology, University of Cambridge) , Stefan Honing (University of Cologne) , David J. Owen (Cambridge Institute for Medical Research (CIMR))
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Developmental Cell , VOL 50 , PAGES 494 - 508.e11

State: Published (Approved)
Published: August 2019

Open Access Open Access

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.

Journal Keywords: clathrin-mediated endocytosis; regulation by phosphorylation; AP2 endocytic adaptor; NECAP; SNX9; AAK1; Numb-associated kinases (NAK); NMRcrystallography; TIRF

Subject Areas: Chemistry, Biology and Bio-materials


Instruments: I02-Macromolecular Crystallography , I04-1-Macromolecular Crystallography (fixed wavelength) , I04-Macromolecular Crystallography

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