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1097-417217422018Jul12CellCellA Flat BAR Protein Promotes Actin Polymerization at the Base of Clathrin-Coated Pits.325337.e14325-337.e1410.1016/j.cell.2018.05.020S0092-8674(18)30597-XMultiple proteins act co-operatively in mammalian clathrin-mediated endocytosis (CME) to generate endocytic vesicles from the plasma membrane. The principles controlling the activation and organization of the actin cytoskeleton during mammalian CME are, however, not fully understood. Here, we show that the protein FCHSD2 is a major activator of actin polymerization during CME. FCHSD2 deletion leads to decreased ligand uptake caused by slowed pit maturation. FCHSD2 is recruited to endocytic pits by the scaffold protein intersectin via an unusual SH3-SH3 interaction. Here, its flat F-BAR domain binds to the planar region of the plasma membrane surrounding the developing pit forming an annulus. When bound to the membrane, FCHSD2 activates actin polymerization by a mechanism that combines oligomerization and recruitment of N-WASP to PI(4,5)P2, thus promoting pit maturation. Our data therefore describe a molecular mechanism for linking spatiotemporally the plasma membrane to a force-generating actin platform guiding endocytic vesicle maturation.Copyright © 2018 MRC Laboratory of Molecular Biology. Published by Elsevier Inc. All rights reserved.Almeida-SouzaLeonardoLMRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK. Electronic address: lalmeida@mrc-lmb.cam.ac.uk.FrankRene A WRAWMRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK.García-NafríaJavierJMRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK.ColussiAdelineAMRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK.GunawardanaNushanNMRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK.JohnsonChristopher MCMMRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK.YuMinminMMRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK.HowardGillianGMRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK.AndrewsByronBMRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK.VallisYvonneYMRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK.McMahonHarvey THTMRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK. Electronic address: hmm@mrc-lmb.cam.ac.uk.engMC_U105178795MRC_Medical Research CouncilUnited KingdomJournal ArticleResearch Support, Non-U.S. Gov't20180607
United StatesCell04130660092-86740Adaptor Proteins, Vesicular Transport0Carrier Proteins0Clathrin0FCHSD2 protein, human0ITSN1 protein, human0Liposomes0Membrane Proteins0RNA, Small Interfering0Wiskott-Aldrich Syndrome Protein, NeuronalIMActin CytoskeletonphysiologyAdaptor Proteins, Vesicular TransportchemistrygeneticsmetabolismCarrier Proteinsantagonists & inhibitorsgeneticsmetabolismCell MembranechemistrymetabolismClathrinmetabolismClathrin-Coated VesiclesmetabolismEndocytosisHeLa CellsHumansLiposomeschemistrymetabolismMembrane Proteinsantagonists & inhibitorsgeneticsmetabolismMicroscopy, FluorescenceModels, MolecularMutagenesis, Site-DirectedRNA InterferenceRNA, Small InterferingmetabolismWiskott-Aldrich Syndrome Protein, Neuronalchemistrymetabolismsrc Homology DomainsARP2/3BAR domainFCHSD2N-WASP activationNervous Wreckactin cytoskeletonclathrin-mediated endocytosiscytoskeletal forcesintersectinmembrane deformation2017572018228201859201861260201944602018612602018712ppublish29887380PMC605726910.1016/j.cell.2018.05.020S0092-8674(18)30597-XBattye T.G.G., Kontogiannis L., Johnson O., Powell H.R., Leslie A.G.W. iMOSFLM: a new graphical interface for diffraction-image processing with MOSFLM. 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